Natural Combination Hormone Replacement Formulations and Therapies

ABSTRACT

Pharmaceutical compositions for co-administering estradiol and progesterone to a human subject in need thereof are provided. In some embodiments, the pharmaceutical composition comprises solubilized estradiol, suspended progesterone, and a solubilizing agent comprising a medium chain (C6-C12) oil.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a divisional of U.S. patent application Ser. No.14/719,933, filed May 22, 2015, which claims priority to U.S.Provisional Application Ser. No. 62/002,090, filed May 22, 2014, thecontent of each of which is incorporated by reference herein in itsentirety.

FIELD OF THE INVENTION

This application relates to pharmaceutical compositions and methods forhormone replacement therapy.

BACKGROUND OF THE INVENTION

Hormone Replacement Therapy (HRT) is a medical treatment that involvesthe use of one or more of a group of medications designed to increasehormone levels in women who lack adequate hormone production. HRT canmitigate and prevent symptoms caused by diminished circulating estrogenand progesterone hormones in a pre-menopausal, peri-menopausal,menopausal or post-menopausal subject.

BRIEF SUMMARY OF THE INVENTION

In one aspect, pharmaceutical compositions for co-administeringestradiol and progesterone to a subject in need of natural hormonereplacement therapies are provided. In some embodiments, thepharmaceutical composition comprises: solubilized estradiol, suspendedprogesterone, and a solubilizing agent, wherein the solubilizing agentis a medium chain (C6-C12) oil and wherein the pharmaceuticalcomposition, when administered to a subject, produces in a plasma samplefrom the subject one or more pharmacokinetic parameters as describedherein (e.g., an area under the curve (AUC)_((0-t)) or a C_(max) forestradiol, progesterone, estrone, or total estrone as described herein,e.g., in Tables 18-21).

In some embodiments, the pharmaceutical composition comprises asolubilizing agent that comprises a glyceride of at least one C6-C12fatty acid. In some embodiments, the glyceride ester is a mixture ofmono- and diglycerides (e.g., glyceryl caprylate/caprate). In someembodiments, the fatty acid is predominantly a C8 to C10 fatty acid. Insome embodiments, the pharmaceutical composition further comprises asurfactant (e.g., lauroyl polyoxyglyceride). In some embodiments, thepharmaceutical composition comprises estradiol at a dosage of about0.05, 0.1, 0.125, 0.15, 0.20, 0.25, 0.30, 0.35, 0.375, 0.40, 0.45, 0.50,0.55, 0.60, 0.625, 0.65, 0.70, 0.75, 0.80, 0.85, 0.90, 0.95, 1.00,1.125, 1.25, 1.375, 1.50, 1.625, 1.75, or 2.00 mg, and comprisesprogesterone at a dosage of about 25, 50, 75, 100, 125, 150, 175, 200,250, 300, 350, or 400 mg. In some embodiments, the pharmaceuticalcomposition comprises estradiol at a dosage of about 0.25 mg andcomprises progesterone at a dosage of about 50 mg. In some embodiments,the pharmaceutical composition comprises estradiol at a dosage of about0.50 mg and comprises progesterone at a dosage of about 50 mg. In someembodiments, the pharmaceutical composition comprises estradiol at adosage of about 0.50 mg and comprises progesterone at a dosage of about100 mg. In some embodiments, the pharmaceutical composition comprisesestradiol at a dosage of about 1 mg and comprises progesterone at adosage of about 100 mg. In some embodiments, the pharmaceuticalcomposition comprises estradiol at a dosage of about 2 mg and comprisesprogesterone at a dosage of about 200 mg.

In some embodiments, the pharmaceutical composition comprises about 0.25mg estradiol and about 50 mg progesterone, and administration of thecomposition to the subject produces, in a plasma sample from thesubject, one or more parameters selected from:

-   -   (i) an area under the curve (AUC)_((0-t)) for estradiol that is        from 140.3733 pg·hr/ml to 219.3333 pg·hr/ml;    -   (ii) a C_(max) for estradiol that is from 6.4790 pg/ml to        10.1235 pg/ml;    -   (iii) an AUC_((0-t)) for progesterone that is from 24.0174        ng·hr/ml to 37.5272 ng·hr/ml; and    -   (iv) a C_(max) for progesterone that is from 17.8444 ng/ml to        27.8819 ng/ml.

In some embodiments, administration of the composition to the subjectfurther produces, in a plasma sample from the subject, one or bothparameters selected from: an AUC_((0-t)) for estrone that is from909.6091 pg·hr/ml to 1421.2642 pg·hr/ml; and a C_(max) for estrone thatis from 42.6549 pg/ml to 66.6483 pg/ml.

In some embodiments, administration of the composition to subjectfurther produces, in a plasma sample from the subject, one or bothparameters selected from: an AUC_((0-t)) for total estrone that is from20.1752 ng·hr/ml to 31.5238 ng·hr/ml; and a C_(max) for total estronethat is from 3.5429 ng/ml to 5.5358 ng/ml.

In some embodiments, the pharmaceutical composition comprises about 0.25mg estradiol and about 50 mg progesterone, and administration of thecomposition to a subject produces, in a plasma sample from the subject,the following parameters:

-   -   (i) one or both of (a) an AUC_((0-t)) for estradiol that is from        140.3733 pg·hr/ml to 219.3333 pg·hr/ml and (b) a C_(max) for        estradiol that is from 6.4790 pg/ml to 10.1235 pg/ml; and    -   (ii) one or both of (a) an AUC_((0-t)) for progesterone that is        from 24.0174 ng·hr/ml to 37.5272 ng·hr/ml and (b) a C_(max) for        progesterone that is from 17.8444 ng/ml to 27.8819 ng/ml; and        optionally    -   (iii) one or both of (a) an AUC_((0-t)) for estrone that is from        909.6091 pg·hr/ml to 1421.2642 pg·hr/ml and (b) a C_(max) for        estrone that is from 42.6549 pg/ml to 66.6483 pg/ml; and        optionally    -   (iv) one or both of (a) an AUC_((0-t)) for total estrone that is        from 20.1752 ng·hr/ml to 31.5238 ng·hr/ml and (b) a C_(max) for        total estrone that is from 3.5429 ng/ml to 5.5358 ng/ml.

In some embodiments, a pharmaceutical composition for co-administeringestradiol and progesterone to a human subject in need thereof comprisesabout 0.50 mg estradiol and about 50 mg progesterone, and administrationof the composition to the subject produces, in a plasma sample from thesubject, one or more parameters selected from:

-   -   (i) an AUC_((0-t)) for estradiol that is from 280.7467 pg·hr/ml        to 438.6667 pg·hr/ml;    -   (ii) a C_(max) for estradiol that is from 12.9580 pg/ml to        20.2469 pg/ml;    -   (iii) an AUC_((0-t)) for progesterone that is from 24.0174        ng·hr/ml to 37.5272 ng·hr/ml; and    -   (iv) a C_(max) for progesterone that is from 17.8444 ng/ml to        27.8819 ng/ml.

In some embodiments, administration of the composition to the subjectfurther produces, in a plasma sample from the subject, one or bothparameters selected from: an AUC_((0-t)) for estrone that is from1819.2181 pg·hr/ml to 2842.5283 pg·hr/ml, and a C_(max) for estrone thatis from 85.3098 pg/ml to 133.2966 pg/ml.

In some embodiments, administration of the composition to the subjectfurther produces, in a plasma sample from the subject, one or bothparameters selected from: an AUC_((0-t)) for total estrone that is from40.3505 ng·hr/ml to 63.0476 ng·hr/ml, and a C_(max) for total estronethat is from 7.0858 ng/ml to 11.0715 ng/ml.

In some embodiments, the pharmaceutical composition comprises about 0.50mg estradiol and about 50 mg progesterone, and administration of thecomposition to a subject produces, in a plasma sample from the subject,the following parameters:

-   -   (i) one or both of (a) an UC_((0-t)) for estradiol that is from        280.7467 pg·hr/ml to 438.6667 pg·hr/ml and (b) a C_(max) for        estradiol that is from 12.9580 pg/ml to 20.2469 pg/ml; and    -   (ii) one or both of (a) an AUC_((0-t)) for progesterone that is        from 24.0174 ng·hr/ml to 37.5272 ng·hr/ml and (b) a C_(max) for        progesterone that is from 17.8444 ng/ml to 27.8819 ng/ml; and        optionally    -   (iii) one or both of (a) an AUC_((0-t)) for estrone that is from        1819.2181 pg·hr/ml to 2842.5283 pg·hr/ml and (b) a C_(max) for        estrone that is from 85.3098 pg/ml to 133.2966 pg/ml; and        optionally    -   (iv) one or both of (a) an AUC_((0-t)) for total estrone that is        from 40.3505 ng·hr/ml to 63.0476 ng·hr/ml and (b) a C_(max) for        total estrone that is from 7.0858 ng/ml to 11.0715 ng/ml.

In some embodiments, a pharmaceutical composition for co-administeringestradiol and progesterone to a human subject in need thereof comprisesabout 0.50 mg estradiol and about 100 mg progesterone, andadministration of the composition to the subject produces, in a plasmasample from the subject, one or more parameters selected from:

-   -   (i) an area under the curve (AUC)_((0-t)) for estradiol that is        from 280.7467 pg·hr/ml to 438.6667 pg·hr/ml;    -   (ii) a C_(max) for estradiol that is from 12.9580 pg/ml to        20.2469 pg/ml;    -   (iii) an AUC_((0-t)) for progesterone that is from 48.0348        ng·hr/ml to 75.0543 ng·hr/ml; and    -   (iv) a C_(max) for progesterone that is from 35.6889 ng/ml to        55.7639 ng/ml.

In some embodiments, administration of the composition to the subjectfurther produces, in a plasma sample from the subject, one or bothparameters selected from: an AUC_((0-t)) for estrone that is from1819.2181 pg·hr/ml to 2842.5283 pg·hr/ml, and a C_(max) for estrone thatis from 85.3098 pg/ml to 133.2966 pg/ml.

In some embodiments, administration of the composition to the subjectfurther produces, in a plasma sample from the subject, one or bothparameters selected from: an AUC_((0-t)) for total estrone that is from40.3505 ng·hr/ml to 63.0476 ng·hr/ml, and a C_(max) for total estronethat is from 7.0858 ng/ml to 11.0715 ng/ml.

In some embodiments, the pharmaceutical composition comprises about 0.50mg estradiol and about 100 mg progesterone, and administration of thecomposition to a subject produces, in a plasma sample from the subject,the following parameters:

-   -   (i) one or both of (a) an AUC_((0-t)) for estradiol that is from        280.7467 pg·hr/ml to 438.6667 pg·hr/ml and (b) a C_(max) for        estradiol that is from 12.9580 pg/ml to 20.2469 pg/ml; and    -   (ii) one or both of (a) an AUC_((0-t)) for progesterone that is        from 48.0348 ng·hr/ml to 75.0543 ng·hr/ml and (b) a C_(max) for        progesterone that is from 35.6889 ng/ml to 55.7639 ng/ml; and        optionally    -   (iii) one or both of (a) an AUC_((0-t)) for estrone that is from        1819.2181 pg·hr/ml to 2842.5283 pg·hr/ml and (b) a C_(max) for        estrone that is from 85.3098 pg/ml to 133.2966 pg/ml; and        optionally    -   (iv) one or both of (a) an AUC_((0-t)) for total estrone that is        from 40.3505 ng·hr/ml to 63.0476 ng·hr/ml and (b) a C_(max) for        total estrone that is from 7.0858 ng/ml to 11.0715 ng/ml.

In some embodiments, a pharmaceutical composition for co-administeringestradiol and progesterone to a human subject in need thereof comprisesabout 1 mg estradiol and about 100 mg progesterone, and administrationof the composition to the subject produces, in a plasma sample from thesubject, one or more parameters selected from:

-   -   (i) an area under the curve (AUC)_((0-t)) for estradiol that is        from 561.4933 pg·hr/ml to 877.3333 pg·hr/ml;    -   (ii) a C_(max) for estradiol that is from 25.9161 pg/ml to        40.4939 pg/ml;    -   (iii) an AUC_((0-t)) for progesterone that is from 48.0348        ng·hr/ml to 75.0543 ng·hr/ml; and    -   (iv) a C_(max) for progesterone that is from 35.6889 ng/ml to        55.7639 ng/ml.

In some embodiments, administration of the composition to the subjectfurther produces, in a plasma sample from the subject, one or bothparameters selected from: an AUC_((0-t)) for estrone that is from3638.4363 pg·hr/ml to 5685.0567 pg·hr/ml, and a C_(max) for estrone thatis from 170.6197 pg/ml to 266.5933 pg/ml.

In some embodiments, administration of the composition to the subjectfurther produces, in a plasma sample from the subject, one or bothparameters selected from: an AUC_((0-t)) for total estrone that is from80.7010 ng·hr/ml to 126.0953 ng·hr/ml, and a C_(max) for total estronethat is from 14.1716 ng/ml to 22.1431 ng/ml.

In some embodiments, the pharmaceutical composition comprises about 0.50mg estradiol and about 100 mg progesterone, and administration of thecomposition to a subject produces, in a plasma sample from the subject,the following parameters:

-   -   (i) one or both of (a) an AUC_((0-t)) for estradiol that is from        561.4933 pg·hr/ml to 877.3333 pg·hr/ml and (b) a C_(max) for        estradiol that is from 25.9161 pg/ml to 40.4939 pg/ml; and    -   (ii) one or both of (a) an AUC_((0-t)) for progesterone that is        from 48.0348 ng·hr/ml to 75.0543 ng·hr/ml and (b) a C_(max) for        progesterone that is from 35.6889 ng/ml to 55.7639 ng/ml; and        optionally    -   (iii) one or both of (a) an AUC_((0-t)) for estrone that is from        3638.4363 pg·hr/ml to 5685.0567 pg·hr/ml and (b) a C_(max) for        estrone that is from 170.6197 pg/ml to 266.5933 pg/ml; and        optionally    -   (iv) one or both of (a) an AUC_((0-t)) for total estrone that is        from 80.7010 ng·hr/ml to 126.0953 ng·hr/ml and (b) a C_(max) for        total estrone that is from 14.1716 ng/ml to 22.1431 ng/ml.

In some embodiments, the pharmaceutical composition has the blood plasmaestradiol concentration profile of FIG. 1. In some embodiments, thepharmaceutical composition has the blood plasma progesteroneconcentration profile of FIG. 2. In some embodiments, the pharmaceuticalcomposition has the blood plasma estrone concentration profile of FIG.3. In some embodiments, the pharmaceutical composition has the bloodplasma total estrone concentration profile of FIG. 4.

In some embodiments, the one or more parameters as described herein(e.g., the AUC_((0-t)) or C_(max) for progesterone, estradiol, estrone,or total estrone) are measured at regular intervals (e.g., about every30 minutes, about every 60 minutes, or about every 90 minutes) or atirregular intervals over a period of time such as 24 hours or 48 hours.In some embodiments, the one or more parameters as described herein(e.g., the AUC_((0-t)) or C_(max) for progesterone, estradiol, estrone,or total estrone) are measured at about 0.25 hr, 0.5 hr, 0.67 hr, 0.83hr, 1 hr, 1.33 hr, 1.67 hr, 2 hr, 2.5 hr, 3 hr, 4 hr, 5 hr, 6 hr, 7 hr,8 hr, 10 hr, 12 hr, 18 hr, 24 hr, 36 hr, or 48 hr after administeringthe pharmaceutical composition to the subject. In some embodiments, theone or more parameters as described herein are measured at regular orirregular intervals following the administration of a single dose or ofa first dose of the pharmaceutical composition to the subject.

In another aspect, methods of treating a subject are provided. In someembodiments, the subject has a condition that is caused at least in partby an estrogen deficiency (e.g., one or more symptoms of menopause, suchas vasomotor symptoms). In some embodiments, the method comprisesadministering to the subject a pharmaceutical composition comprisingsolubilized estradiol, suspended progesterone, and a solubilizing agentthat comprises a medium chain (C6-C12) oil as described herein, whereinadministration of the pharmaceutical composition produces, in a plasmasample from the subject, one or more pharmacokinetic parameters asdescribed herein. In some embodiments, the method comprisesadministering a pharmaceutical composition comprising estradiol at adosage of about 0.05, 0.1, 0.125, 0.15, 0.20, 0.25, 0.30, 0.35, 0.375,0.40, 0.45, 0.50, 0.55, 0.60, 0.625, 0.65, 0.70, 0.75, 0.80, 0.85, 0.90,0.95, 1.00, 1.125, 1.25, 1.375, 1.50, 1.625, 1.75, or 2.00 mg, andcomprising progesterone at a dosage of about 25, 50, 75, 100, 125, 150,175, 200, 250, 300, 350, or 400 mg. In some embodiments, the methodcomprises administering a pharmaceutical composition comprising:estradiol at a dosage of about 0.25 mg and progesterone at a dosage ofabout 50 mg; estradiol at a dosage of about 0.50 mg and progesterone ata dosage of about 50 mg; estradiol at a dosage of about 0.50 mg andprogesterone at a dosage of about 100 mg; estradiol at a dosage of about1 mg and progesterone at a dosage of about 100 mg; or estradiol at adosage of about 2 mg and progesterone at a dosage of about 200 mg.

In some embodiments, the method comprises administering to the subject apharmaceutical composition comprising about 0.25 mg estradiol and about50 mg progesterone, wherein administration of the pharmaceuticalcomposition produces, in a plasma sample from the subject, one or moreparameters selected from:

-   -   (i) an area under the curve (AUC)_((0-t)) for estradiol that is        from 140.3733 pg·hr/ml to 219.3333 pg·hr/ml;    -   (ii) a C_(max) for estradiol that is from 6.4790 pg/ml to        10.1235 pg/ml;    -   (iii) an AUC_((0-t)) for progesterone that is from 24.0174        ng·hr/ml to 37.5272 ng·hr/ml; and    -   (iv) a C_(max) for progesterone that is from 17.8444 ng/ml to        27.8819 ng/ml.

In some embodiments, administration of the pharmaceutical compositionfurther produces, in a plasma sample from the subject, one or moreparameters selected from: an AUC_((0-t)) for estrone that is from909.6091 pg·hr/ml to 1421.2642 pg·hr/ml; a C_(max) for estrone that isfrom 42.6549 pg/ml to 66.6483 pg/ml; an AUC_((0-t)) for total estronethat is from 20.1752 ng·hr/ml to 31.5238 ng·hr/ml; and a C_(max) fortotal estrone that is from 3.5429 ng/ml to 5.5358 ng/ml.

In some embodiments, the method comprises administering to the subject apharmaceutical composition comprising about 0.25 mg estradiol and about50 mg progesterone, wherein administration of the pharmaceuticalcomposition produces, in a plasma sample from the subject, the followingparameters:

-   -   (i) one or both of (a) an AUC_((0-t)) for estradiol that is from        140.3733 pg·hr/ml to 219.3333 pg·hr/ml and (b) a C_(max) for        estradiol that is from 6.4790 pg/ml to 10.1235 pg/ml; and    -   (ii) one or both of (a) an AUC_((0-t)) for progesterone that is        from 24.0174 ng·hr/ml to 37.5272 ng·hr/ml and (b) a C_(max) for        progesterone that is from 17.8444 ng/ml to 27.8819 ng/ml; and        optionally    -   (iii) one or both of (a) an AUC_((0-t)) for estrone that is from        909.6091 pg·hr/ml to 1421.2642 pg·hr/ml and (b) a C_(max) for        estrone that is from 42.6549 pg/ml to 66.6483 pg/ml; and        optionally    -   (iv) one or both of (a) an an AUC_((0-t)) for total estrone that        is from 20.1752 ng·hr/ml to 31.5238 ng·hr/ml and (b) a C_(max)        for total estrone that is from 3.5429 ng/ml to 5.5358 ng/ml.

In some embodiments, the method comprises administering to the subject apharmaceutical composition comprising about 0.50 mg estradiol and about50 mg progesterone, wherein administration of the pharmaceuticalcomposition produces, in a plasma sample from the subject, one or moreparameters selected from:

-   -   (i) an AUC_((0-t)) for estradiol that is from 280.7467 pg·hr/ml        to 438.6667 pg·hr/ml;    -   (ii) a C_(max) for estradiol that is from 12.9580 pg/ml to        20.2469 pg/ml;    -   (iii) an AUC_((0-t)) for progesterone that is from 24.0174        ng·hr/ml to 37.5272 ng·hr/ml; and    -   (iv) a C_(max) for progesterone that is from 17.8444 ng/ml to        27.8819 ng/ml.

In some embodiments, administration of the composition to the subjectfurther produces, in a plasma sample from the subject, one or moreparameters selected from: an AUC_((0-t)) for estrone that is from1819.2181 pg·hr/ml to 2842.5283 pg·hr/ml; a C_(max) for estrone that isfrom 85.3098 pg/ml to 133.2966 pg/ml; an AUC_((0-t)) for total estronethat is from 40.3505 ng·hr/ml to 63.0476 ng·hr/ml; and a C_(max) fortotal estrone that is from 7.0858 ng/ml to 11.0715 ng/ml.

In some embodiments, the method comprises administering to the subject apharmaceutical composition comprising about 0.50 mg estradiol and about50 mg progesterone, wherein administration of the pharmaceuticalcomposition produces, in a plasma sample from the subject, the followingparameters:

-   -   (i) one or both of (a) an AUC_((0-t)) for estradiol that is from        280.7467 pg·hr/ml to 438.6667 pg·hr/ml and (b) a C_(max) for        estradiol that is from 12.9580 pg/ml to 20.2469 pg/ml; and    -   (ii) one or both of (a) an AUC_((0-t)) for progesterone that is        from 24.0174 ng·hr/ml to 37.5272 ng·hr/ml and (b) a C_(max) for        progesterone that is from 17.8444 ng/ml to 27.8819 ng/ml; and        optionally    -   (iii) one or both of (a) an AUC_((0-t)) for estrone that is from        1819.2181 pg·hr/ml to 2842.5283 pg·hr/ml and (b) a C_(max) for        estrone that is from 85.3098 pg/ml to 133.2966 pg/ml; and        optionally    -   (iv) one or both of (a) an an AUC_((0-t)) for total estrone that        is from 40.3505 ng·hr/ml to 63.0476 ng·hr/ml and (b) a C_(max)        for total estrone that is from 7.0858 ng/ml to 11.0715 ng/ml.

In some embodiments, the method comprises administering to the subject apharmaceutical composition comprising about 0.50 mg estradiol and about100 mg progesterone, wherein administration of the pharmaceuticalcomposition produces, in a plasma sample from the subject, one or moreparameters selected from:

-   -   (i) an area under the curve (AUC)_((0-t)) for estradiol that is        from 280.7467 pg·hr/ml to 438.6667 pg·hr/ml;    -   (ii) a C_(max) for estradiol that is from 12.9580 pg/ml to        20.2469 pg/ml;    -   (iii) an AUC_((0-t)) for progesterone that is from 48.0348        ng·hr/ml to 75.0543 ng·hr/ml; and    -   (iv) a C_(max) for progesterone that is from 35.6889 ng/ml to        55.7639 ng/ml.

In some embodiments, administration of the composition to the subjectfurther produces, in a plasma sample from the subject, one or moreparameters selected from: an AUC_((0-t)) for estrone that is from1819.2181 pg·hr/ml to 2842.5283 pg·hr/ml; a C_(max) for estrone that isfrom 85.3098 pg/ml to 133.2966 pg/ml; an AUC_((0-t)) for total estronethat is from 40.3505 ng·hr/ml to 63.0476 ng·hr/ml, and a C_(max) fortotal estrone that is from 7.0858 ng/ml to 11.0715 ng/ml.

In some embodiments, the method comprises administering to the subject apharmaceutical composition comprising about 0.50 mg estradiol and about100 mg progesterone, wherein administration of the pharmaceuticalcomposition produces, in a plasma sample from the subject, the followingparameters:

-   -   (i) one or both of (a) an AUC_((0-t)) for estradiol that is from        280.7467 pg·hr/ml to 438.6667 pg·hr/ml and (b) a C_(max) for        estradiol that is from 12.9580 pg/ml to 20.2469 pg/ml; and    -   (ii) one or both of (a) an AUC_((0-t)) for progesterone that is        from 48.0348 ng·hr/ml to 75.0543 ng·hr/ml and (b) a C_(max) for        progesterone that is from 35.6889 ng/ml to 55.7639 ng/ml; and        optionally    -   (iii) one or both of (a) an AUC_((0-t)) for estrone that is from        1819.2181 pg·hr/ml to 2842.5283 pg·hr/ml and (b) a C_(max) for        estrone that is from 85.3098 pg/ml to 133.2966 pg/ml; and        optionally    -   (iv) one or both of (a) AUC_((0-t)) for total estrone that is        from 40.3505 ng·hr/ml to 63.0476 ng·hr/ml and (b) a C_(max) for        total estrone that is from 7.0858 ng/ml to 11.0715 ng/ml.

In some embodiments, the method comprises administering to the subject apharmaceutical composition comprising about 1 mg estradiol and about 100mg progesterone, wherein administration of the pharmaceuticalcomposition produces, in a plasma sample from the subject, one or moreparameters selected from:

-   -   (i) an area under the curve (AUC)_((0-t)) for estradiol that is        from 561.4933 pg·hr/ml to 877.3333 pg·hr/ml;    -   (ii) a C_(max) for estradiol that is from 25.9161 pg/ml to        40.4939 pg/ml;    -   (iii) an AUC_((0-t)) for progesterone that is from 48.0348        ng·hr/ml to 75.0543 ng·hr/ml; and    -   (iv) a C_(max) for progesterone that is from 35.6889 ng/ml to        55.7639 ng/ml.

In some embodiments, administration of the composition to the subjectfurther produces, in a plasma sample from the subject, one or moreparameters selected from: an AUC_((0-t)) for estrone that is from3638.4363 pg·hr/ml to 5685.0567 pg·hr/ml; a C_(max) for estrone that isfrom 170.6197 pg/ml to 266.5933 pg/ml; an AUC_((0-t)) for total estronethat is from 80.7010 ng·hr/ml to 126.0953 ng·hr/ml; and a C_(max) fortotal estrone that is from 14.1716 ng/ml to 22.1431 ng/ml.

In some embodiments, the method comprises administering to the subject apharmaceutical composition comprising about 1 mg estradiol and about 100mg progesterone, wherein administration of the pharmaceuticalcomposition produces, in a plasma sample from the subject, the followingparameters:

-   -   (i) one or both of (a) an AUC_((0-t)) for estradiol that is from        561.4933 pg·hr/ml to 877.3333 pg·hr/ml and (b) a C_(max) for        estradiol that is from 25.9161 pg/ml to 40.4939 pg/ml; and    -   (ii) one or both of (a) an AUC_((0-t)) for progesterone that is        from 48.0348 ng·hr/ml to 75.0543 ng·hr/ml and (b) a C_(max) for        progesterone that is from 35.6889 ng/ml to 55.7639 ng/ml; and        optionally    -   (iii) one or both of (a) an AUC_((0-t)) for estrone that is from        3638.4363 pg·hr/ml to 5685.0567 pg·hr/ml and (b) a C_(max) for        estrone that is from 170.6197 pg/ml to 266.5933 pg/ml; and        optionally    -   (iv) one or both of (a) an AUC_((0-t)) for total estrone that is        from 80.7010 ng·hr/ml to 126.0953 ng·hr/ml and (b) a C_(max) for        total estrone that is from 14.1716 ng/ml to 22.1431 ng/ml.

In still another aspect, pharmaceutical compositions for use in a methodof treating a disease or condition that is caused at least in part by anestrogen deficiency are provided. In some embodiments, thepharmaceutical composition comprises solubilized estradiol, suspendedprogesterone, and a solubilizing agent that comprises a medium chain(C6-C12) oil, wherein the treatment produces, in a plasma sample fromthe subject, one or more pharmacokinetic parameters as described herein(e.g., an AUC_((0-t)) or C_(max) for estradiol, progesterone, estrone,or total estrone as described herein, e.g., as described in any ofTables 18-21). In some embodiments, the pharmaceutical compositions foruse in a method of treating a disease or condition that is caused atleast in part by an estrogen deficiency comprise estradiol at a dosageof about 0.05, 0.1, 0.125, 0.15, 0.20, 0.25, 0.30, 0.35, 0.375, 0.40,0.45, 0.50, 0.55, 0.60, 0.625, 0.65, 0.70, 0.75, 0.80, 0.85, 0.90, 0.95,1.00, 1.125, 1.25, 1.375, 1.50, 1.625, 1.75, or 2.00 mg, and compriseprogesterone at a dosage of about 25, 50, 75, 100, 125, 150, 175, 200,250, 300, 350, or 400 mg.

In some embodiments, a pharmaceutical composition for use in a method oftreating a disease or condition that is caused at least in part by anestrogen deficiency (e.g., one or more symptoms of menopause) comprisesestradiol at a dosage of about 0.25 mg and progesterone at a dosage ofabout 50 mg, and produces one or more pharmacokinetic values disclosedin Table 18 following administration of a single dose of thepharmaceutical composition to a subject (e.g., about 24 hours or about48 hours after administration).

In some embodiments, a pharmaceutical composition for use in a method oftreating a disease or condition that is caused at least in part by anestrogen deficiency (e.g., one or more symptoms of menopause) comprisesestradiol at a dosage of about 0.50 mg and progesterone at a dosage ofabout 50 mg, and produces one or more pharmacokinetic values disclosedin Table 19 following administration of a single dose of thepharmaceutical composition to a subject (e.g., about 24 hours or about48 hours after administration).

In some embodiments, a pharmaceutical composition for use in a method oftreating a disease or condition that is caused at least in part by anestrogen deficiency (e.g., one or more symptoms of menopause) comprisesestradiol at a dosage of about 0.50 mg and progesterone at a dosage ofabout 100 mg, and produces one or more pharmacokinetic values disclosedin Table 20 following administration of a single dose of thepharmaceutical composition to a subject (e.g., about 24 hours or about48 hours after administration).

In some embodiments, a pharmaceutical composition for use in a method oftreating a disease or condition that is caused at least in part by anestrogen deficiency (e.g., one or more symptoms of menopause) comprisesestradiol at a dosage of about 1 mg and progesterone at a dosage ofabout 100 mg, and produces one or more pharmacokinetic values disclosedin Table 21 following administration of a single dose of thepharmaceutical composition to a subject (e.g., about 24 hours or about48 hours after administration).

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 illustrates a semilogarithmic plot of mean plasma concentration(pg/ml) over time (hrs) for estradiol.

FIG. 2 illustrates a semilogarithmic plot of mean plasma concentration(ng/ml) over time (hrs) for progesterone.

FIG. 3 illustrates a semilogarithmic plot of mean plasma concentration(pg/ml) over time (hrs) for estrone.

FIG. 4 illustrates a semilogarithmic plot of mean plasma concentration(ng/ml) over time (hrs) for total estrone.

DETAILED DESCRIPTION OF THE INVENTION

In the following detailed description of embodiments of this disclosure,reference is made to the accompanying drawings in which like referencesindicate similar elements, and in which is shown, by way ofillustration, specific embodiments in which this disclosure may bepracticed. These embodiments are described in sufficient detail toenable those skilled in the art to practice this disclosure, and it isto be understood that other embodiments may be utilized and that otherchanges may be made without departing from the scope of this disclosure.The following detailed description is, therefore, not to be taken in alimiting sense, and the scope of this disclosure is defined only by theappended claims. As used in this disclosure, the term “or” shall beunderstood to be defined as a logical disjunction (i.e., and/or) andshall not indicate an exclusive disjunction unless expressly indicatedas such with the term “either,” “unless,” “alternatively,” and words ofsimilar effect.

I. DEFINITIONS

The term “area under the curve” (“AUC”) refers to the area under thecurve defined by changes in the blood, plasma, or serum concentration ofan active pharmaceutical ingredient (e.g., estradiol or progesterone),or one or more metabolites of the active pharmaceutical ingredient, overtime following the administration of a dose of the active pharmaceuticalingredient. “AUC_(0-∞).” is the area under the concentration-time curveextrapolated to infinity following the administration of a dose.“AUC_(0-t)” is the area under the concentration-time curve from timezero to time t following the administration of a dose, wherein t is thelast time point with measurable concentration.

The term “C_(max)” refers to the maximum value of blood, plasma, orserum concentration shown on the curve that represents changes in blood,plasma, or serum concentrations of an active pharmaceutical ingredient(e.g., progesterone or estradiol), or one or more metabolites of theactive pharmaceutical ingredient, over time.

The term “T_(max)” refers to the time that it takes for the blood,plasma, or serum concentration of an active pharmaceutical ingredient(e.g., estradiol or progesterone), or of one or more metabolites of theactive pharmaceutical ingredient, to reach the maximum value.

Collectively, AUC, C_(max), and, optionally, T_(max) are the principalpharmacokinetic parameters that can characterize the pharmacokineticresponse of a particular drug product, such as progesterone orestradiol, in an animal, especially a mammal, including human, subject.

An “active pharmaceutical ingredient” (API), as used herein, means theactive compound or compounds used in formulating a drug product. APIsare generally safe for administering to animals, especially mammals,including humans, according to established governmental standards,including those promulgated by the United States Food and DrugAdministration.

The term “bioavailability” has the meaning as defined in 21 C.F.R. §320.1(a): the rate and extent to which an API or active ingredient oractive moiety is absorbed from a drug product and becomes available atthe site of action. For drug products that are not intended to beabsorbed into the bloodstream, bioavailability may be assessed bymeasurements intended to reflect the rate and extent to which the API oractive ingredient or active moiety becomes available at the site ofaction. For example, bioavailability can be measured as the amount ofAPI in the blood (whole blood, serum, or plasma) as a function of time.In embodiments, the amount of API is measured in blood plasma.Pharmacokinetic (PK) parameters such as AUC, C_(max), or T_(max) may beused to measure and assess bioavailability.

The term “bioequivalent” has the meaning as defined in 21 C.F.R. §320.1(e): the absence of a significant difference in the rate and extentto which the API or active ingredient or active moiety in pharmaceuticalequivalents or pharmaceutical alternatives becomes available at the siteof drug action when administered at the same molar dose under similarconditions in an appropriately designed study. Where there is anintentional difference in rate (e.g., in certain extended release dosageforms or modified release dosage forms), certain pharmaceuticalequivalents or alternatives may be considered bioequivalent if there isno significant difference in the extent to which the active ingredientor moiety from each product becomes available at the site of drugaction. This applies only if the difference in the rate at which theactive ingredient or moiety becomes available at the site of drug actionis intentional and is reflected in the proposed labeling, is notessential to the attainment of effective body drug concentrations onchronic use, and is considered medically insignificant for the drug. Inpractice, two products are considered bioequivalent if the 90%confidence interval of the AUC, C_(max), or optionally T_(max) is within80.00% to 125.00%.

The term “bio-identical hormone” or “body-identical hormone” refers toan active pharmaceutical ingredient that is structurally identical to ahormone naturally or endogenously found in the human body (e.g.,estradiol and progesterone).

The term “estrogen” refers to a group of several female sex hormonesproduced primarily by the ovaries, including estradiol, estrone, andestriol. As used herein, unless otherwise specified, estrogen refers toestradiol.

The term “estradiol” refers to (17β)-estra-1,3,5(10)-triene-3,17-diol.Estradiol is also interchangeably called 17β-estradiol, oestradiol, orE2, and is found endogenously in the human body. As used herein,estradiol refers to the bio-identical or body-identical form ofestradiol found in the human body having the structure:

As used herein, unless specified, estradiol includes estradiol inanhydrous or hemihydrate forms. For the purposes of this disclosure, theanhydrous form or the hemihydrate form can be substituted for the otherby accounting for the water or lack of water according to well-known andunderstood techniques.

The term “solubilized estradiol” means that the estradiol or a portionthereof is solubilized or dissolved in the solubilizing agents or theformulations disclosed herein. Solubilized estradiol may includeestradiol that is about 80% solubilized, about 85% solubilized, about90% solubilized, about 95% solubilized, about 96% solubilized, about 97%solubilized, about 98% solubilized, about 99% solubilized or about 100%solubilized. In some embodiments, the estradiol is “fully solubilized”with all or substantially all of the estradiol being solubilized ordissolved in the solubilizing agent. Fully solubilized estradiol mayinclude estradiol that is about 97% solubilized, about 98% solubilized,about 99% solubilized or about 100% solubilized. Solubility can beexpressed as a mass fraction (% w/w, which is also referred to as wt %).

The term “progesterone” refers to pregn-4-ene-3,20-dione. Progesteroneis also interchangeably called P4 and is found endogenously in the humanbody. As used herein, progesterone refers to the bio-identical orbody-identical form of progesterone found in the human body having thestructure:

The term “solubilized progesterone” means that the progesterone or aportion thereof is solubilized or dissolved in the solubilizing agentsor the formulations disclosed herein disclosed herein. In someembodiments, the progesterone is “partially solubilized” with a portionof the progesterone being solubilized or dissolved in the solubilizingagent and a portion of the progesterone being suspended in thesolubilizing agent. Partially solubilized progesterone may includeprogesterone that is about 1% solubilized, about 5% solubilized, about10% solubilized, about 15% solubilized, about 20% solubilized, about 30%solubilized, about 40% solubilized, about 50% solubilized, about 60%solubilized, about 70% solubilized, about 80% solubilized, about 85%solubilized, about 90% solubilized or about 95% solubilized. In otherembodiments, the progesterone is “fully solubilized” with all orsubstantially all of the progesterone being solubilized or dissolved inthe solubilizing agent. Fully solubilized progesterone may includeprogesterone that is about 97% solubilized, about 98% solubilized, about99% solubilized or about 100% solubilized. Solubility can be expressedas a mass fraction (% w/w, which is also referred to as wt %).

The terms “micronized progesterone” and “micronized estradiol,” as usedherein, include micronized progesterone and micronized estradiol,respectively, having an X50 particle size value below about 15 micronsor having an X90 particle size value below about 25 microns.

The term “X50” means that one-half of the particles in a sample aresmaller in diameter than a given number. For example, micronizedprogesterone having an X50 of 5 microns means that, for a given sampleof micronized progesterone, one-half of the particles have a diameter ofless than 5 microns. Similarly, the term “X90” means that ninety percent(90%) of the particles in a sample are smaller in diameter than a givennumber.

The term “solubilizing agent” refers to an agent or combination ofagents that solubilize an active pharmaceutical ingredient (e.g.,estradiol or progesterone). For example and without limitation, suitablesolubilizing agents include medium chain oils and other solvents andco-solvents that solubilize or dissolve an active pharmaceuticalingredient to a desirable extent. Solubilizing agents suitable for usein the formulations disclosed herein are pharmaceutical gradesolubilizing agents (e.g., pharmaceutical grade medium chain oils). Itwill be understood by those of skill in the art that other excipients orcomponents can be added to or mixed with the solubilizing agent toenhance the properties or performance of the solubilizing agent orresulting formulation. Examples of such excipients include, but are notlimited to, surfactants, emulsifiers, thickeners, colorants, flavoringagents, etc. In some embodiments, the solubilizing agent is a mediumchain oil and, in some other embodiments, the medium chain oil iscombined with a co-solvent(s) or other excipient(s).

The term “medium chain” is used to describe the aliphatic chain lengthof fatty acid containing molecules. “Medium chain” specifically refersto fatty acids, fatty acid esters, or fatty acid derivatives thatcontain fatty acid aliphatic tails or carbon chains that contain between6 (C6) and 14 (C14) carbon atoms.

The terms “medium chain fatty acid” and “medium chain fatty acidderivative” are used to describe fatty acids or fatty acid derivativeswith aliphatic tails (i.e., carbon chains) having 6 to 14 carbons. Fattyacids consist of an unbranched aliphatic tail attached to a carboxylicacid functional group. Fatty acid derivatives include, for example,fatty acid esters and fatty acid containing molecules, including,without limitation, mono-, di- and triglycerides that include componentsderived from fatty acids as well as fatty acid esters of ethylene orpropylene glycol. Those of skill will appreciate that the aliphatictails can be saturated or unsaturated (one or more double bonds betweencarbon atoms). In some embodiments, the aliphatic tails are saturated(i.e., no double bonds between carbon atoms). Medium chain fatty acidsor medium chain fatty acid derivatives include those with aliphatictails having 6-14 carbons, including those that are C6-C14, C6-C12,C8-C14, C8-C12, C6-C10, C8-C10, or others. In embodiments, medium chainfatty acids or medium chain fatty acid derivatives are those that aresaturated. Examples include, without limitation, caproic acid, caprylicacid, capric acid, lauric acid, myristic acid, and derivatives thereof.

The term “oil,” as used herein, refers to any pharmaceuticallyacceptable oil, and specifically excluding peanut oil, that can suspendor solubilize any suitable progesterone or estradiol, starting material,or precursor, including micronized progesterone or estradiol asdescribed herein.

The term “medium chain oil” refers to an oil wherein the composition ofthe fatty acid fraction of the oil is substantially medium chain (i.e.,C6 to C14) fatty acids, i.e., the composition profile of fatty acids inthe oil is substantially medium chain. As used herein, “substantially”means that between 20% and 100% (inclusive of the upper and lowerlimits) of the fatty acid fraction of the oil is made up of medium chainfatty acids, i.e., fatty acids with aliphatic tails (i.e., carbonchains) having 6 to 14 carbons. In some embodiments, about 25%, about30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%,about 65%, about 70%, about 75%, about 85%, about 90% or about 95% ofthe fatty acid fraction of the oil is made up of medium chain fattyacids. Those of skill in the art that will readily appreciate that theterms “alkyl content” or “alkyl distribution” of an oil can be used inplace of the term “fatty acid fraction” of an oil in characterizing agiven oil or solubilizing agent, and these terms are usedinterchangeable herein. As such, medium chain oils suitable for use inthe formulations disclosed herein include medium chain oils wherein thefatty acid fraction of the oil is substantially medium chain fattyacids, or medium chain oils wherein the alkyl content or alkyldistribution of the oil is substantially medium chain alkyls (C6-C12alkyls). It will be understood by those of skill in the art that themedium chain oils suitable for use in the formulations disclosed hereinare pharmaceutical grade (e.g., pharmaceutical grade medium chain oils).Examples of medium chain oils include, for example and withoutlimitation, medium chain fatty acids, medium chain fatty acid esters ofglycerol (e.g., for example, mono-, di-, and triglycerides), mediumchain fatty acid esters of propylene glycol, medium chain fatty acidderivatives of polyethylene glycol, and combinations thereof.

The term “ECN” or “equivalent carbon number” means the sum of the numberof carbon atoms in the fatty acid chains of an oil, and can be used tocharacterize an oil as, for example, a medium chain oil or a long-chainoil. For example, tripalmitin (tripalmitic glycerol), which is a simpletriglyceride containing three fatty acid chains of 16 carbon atoms, hasan ECN of 3×16=48. Conversely, a triglyceride with an ECN=40 may have“mixed” fatty acid chain lengths of 8, 16, and 16; 10, 14, and 16; 8,14, and 18; etc. Naturally occurring oils are frequently “mixed” withrespect to specific fatty acids, but tend not to contain both long chainfatty acids and medium chain fatty acids in the same glycerol backbone.Thus, triglycerides with ECNs of 21-42 typically contain predominantlymedium chain fatty acids; while triglycerides with ECNs of greater than43 typically contain predominantly long chain fatty acids. For example,the ECN of corn oil triglyceride in the US Pharmacopeia (USP) would bein the range of 51-54. Medium chain diglycerides with ECNs of 12-28 willoften contain predominantly medium chain fatty acids, while diglycerideswith ECNs of 32 or greater will typically contain predominantly longchain fatty acids. Monoglycerides will have an ECN that matches thechain length of the sole fatty acid chain. Thus, monoglyceride ECNs inthe range of 6-14 contain mainly medium chain fatty acids, andmonoglycerides with ECNs 16 or greater will contain mainly long chainfatty acids.

The average ECN of a medium chain triglyceride oil is typically 21-42.For example, as listed in the USP, medium chain triglycerides have thefollowing composition as the exemplary oil set forth in the table below:

Fatty Acid Tail Length % of Oil Exemplary Oil 6 <2.0 2.0 8 50.0-80.070.0 10 20.0-50.0 25.0 12 <3.0 2.0 14 <1.0 1.0and would have an average ECN of3*[(6*0.02)+(8*0.070)+(10*0.25)+(12*0.02)+(14*0.01)]=25.8. The ECN ofthe exemplary medium chain triglycerides oil can also be expressed as arange (per the ranges set forth in the USP) of 24.9-27.0. For oils thathave mixed mono-, di-, and triglycerides, or single and double fattyacid glycols, the ECN of the entire oil can be determined by calculatingthe ECN of each individual component (e.g., C8 monoglycerides, C8diglycerides, C10 monoglycerides, and C10 diglycerides) and taking thesum of the relative percentage of the component multiplied by the ECNnormalized to a monoglyceride for each component. For example, the oilhaving C8 and C10 mono- and diglycerides shown in the table below has anECN of 8.3, and is thus a medium chain oil:

ECN as % of Oil ECN as % of Oil Fatty Acid Chain [(chain length) ×Normalized to Length % of Oil (% in oil)] Monoglyceride C8 monoglyceride47 8 × 0.47 = 3.76  3.76 C10 monoglyceride 8 10 × 0.08 = 0.8 0.8 C8diglyceride 38 2 × (8 × 0.38) = 6.08/2 = 3.04 6.08 C10 diglyceride 7 2 ×(10 × 0.07) = 1.4/2 = 0.7 1.4 OIL ECN 8.3 (normalized to monoglycerides)Expressed differently, ECN can be calculated as each chain length in thecomposition multiplied by its relative percentage in the oil:(8*0.85)+(10*0.15)=8.3.

The term “excipients,” as used herein, refers to non-activepharmaceutical ingredients such as solubilizing agents, anti-oxidants,oils, lubricants, and others used in formulating pharmaceuticalproducts.

The terms “treat,” “treating,” and “treatment” refer to any indicia ofsuccess in the treatment or amelioration of an injury, disease, orcondition, including any objective or subjective parameter such asabatement; remission; diminishing of symptoms or making the injury,disease, or condition more tolerable to the patient; slowing in the rateof degeneration or decline; or improving a patient's physical or mentalwell-being. The treatment or amelioration of symptoms can be based onobjective or subject parameters, including the results of a physicalexamination, neuropsychiatric examinations, or psychiatric evaluation.

II. PHARMACEUTICAL COMPOSITIONS

In one aspect, this disclosure relates to pharmaceutical compositionsfor co-administering estradiol and progesterone to a human subject inneed thereof. In some embodiments, the composition comprises estradiol,progesterone, and a solubilizing agent (e.g., a medium chain oil, e.g.,a C6-C12 oil). In some embodiments, a pharmaceutical compositioncomprising estradiol, progesterone, and a solubilizing agent asdescribed herein, when administered to a subject or a population ofsubjects, produces one or more AUC, C_(max), or T_(max) parameters forestradiol, progesterone, estrone, or total estrone as described below.

Formulations of Estradiol and Progesterone Compositions

In some embodiments, a pharmaceutical composition for use as describedherein comprises solubilized estradiol with suspended progesterone;solubilized estradiol with both partially solubilized progesterone andpartially suspended progesterone; or solubilized estradiol with fullysolubilized progesterone. In some embodiments, the composition comprisessolubilized estradiol and suspended progesterone. The underlyingformulation concepts provided herein may be used with other natural orsynthetic forms of estradiol and progesterone, although the natural orbio-identical forms of estradiol and progesterone are preferred.

In some embodiments, the composition comprises estradiol at a dosage ofabout 0.05, 0.1, 0.125, 0.15, 0.20, 0.25, 0.30, 0.35, 0.375, 0.40, 0.45,0.50, 0.55, 0.60, 0.625, 0.65, 0.70, 0.75, 0.80, 0.85, 0.90, 0.95, 1.00,1.125, 1.25, 1.375, 1.50, 1.625, 1.75, or 2.00 mg. In some embodiments,the composition comprises progesterone at a dosage of about 25, 50, 75,100, 125, 150, 175, 200, 250, 300, 350, or 400 mg.

In some embodiments, estradiol is solubilized. Solubilized estradiol mayinclude estradiol that is approximately 80% to 100% soluble in asolubilizing agent, including specifically embodiments that are: 80%,81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%,95%, 96%, 97%, 98%, 99%, or 100% soluble in a solubilizing agent.Solubility may be expressed as a mass fraction (% w/w, also referred toas wt %). In some embodiments, estradiol is micronized. In someembodiments, micronized estradiol has an X50 particle size value of lessthan about 15 microns, less than about 10 microns, less than about 5microns or less than about 3 microns. In some embodiments, micronizedestradiol has an X90 particle size value of less than about 25 microns,less than about 20 microns, or less than about 15 microns. In someembodiments, the composition comprises micronized and partiallysolubilized estradiol.

In some embodiments, the composition comprises micronized progesterone.The progesterone active pharmaceutical ingredient may be micronized viaany one of the multiple methods typically utilized by the ordinarilyskilled artisan. In various embodiments, micronized progesterone has anX50 particle size value of less than about 15 microns, less than about10 microns, less than about 5 microns or less than about 3 microns. Invarious embodiments, micronized progesterone has an X90 particle sizevalue of less than about 25 microns, less than about 20 microns, or lessthan about 15 microns. Particle size may be determined in any suitablemanner. For example, a Beckman Coulter LS 13 320 Laser DiffractionParticle Size Analyzer (the “Beckman Device”) may be used to determineparticle size.

Estradiol and progesterone compositions and methods of preparing suchcompositions are described in U.S. Pat. No. 8,633,178; U.S. PublicationNo. 2013/0129818; U.S. Publication No. 2013/0338123; InternationalPublication No. WO 2013/078422; and International Publication No. WO2013/192251; each of which is incorporated by reference in its entirety.

Solubilizing Agents

Estradiol and progesterone compositions of the present disclosure areprepared via blending with a solubilizing agent. In some embodiments,the solubilizing agent is a pharmaceutically acceptable oil thatcomprises a medium chain oil. In some embodiments, the solubilizingagent is a medium chain oil comprised substantially of C6-C12 mediumchains, e.g., at least 20%, at least 30%, at least 40%, at least 50%, atleast 60%, at least 70%, at least 80%, or at least 90% of the chainspresent in the oil are C6-C12. In some embodiments, the oil comprises atleast one medium chain fatty acid such as medium chain fatty acidshaving at least one mono-, di-, or triglyceride, or derivatives thereof,or combinations thereof. In some embodiments, the medium chain oilcomprises at least one medium chain fatty acid or propylene glycol,polyethylene glycol, or glyceride having esters of medium chain fattyacids. In some embodiments, the solubilizing agent is not peanut oil.

In some embodiments, oils used to solubilize estradiol and to suspend,partially suspend and partially solubilize, or fully solubilizeprogesterone include medium chain fatty acid esters, (e.g., esters ofglycerol, polyethylene glycol, or propylene glycol) and mixturesthereof. In some embodiments, the medium chain fatty acids are C6, C8,C10, C12, C6-C12, C8-C12, C6-C10, C8-C10, or C10-C12 fatty acids. Insome embodiments, the medium chain fatty acids are saturated, orpredominantly saturated, e.g., greater than about 50% saturated, greaterthan about 60% saturated, or greater than about 75% saturated. In someembodiments, a solubilizing agent comprises predominantly medium chainlength, saturated fatty acids or derivatives thereof, specificallypredominantly C8 to C12 saturated fatty acids or derivatives thereof.

In some embodiments, medium chain solubilizing agents include, forexample and without limitation, saturated medium chain fatty acids orderivatives of saturated medium chain fatty acids: caproic acid (C6),enanthic acid (C7), caprylic acid (C8), pelargonic acid (C9), capricacid (C10), undecylic acid (C11), lauric acid (C12), tridecylic acid(C13), or myristic acid (C14). In some embodiments, the solubilizingagent comprises oils made of these free medium chain fatty acids, oilsof medium chain fatty acid esters of glycerin, propylene glycol, orethylene glycol, or combinations thereof. These examples comprisepredominantly saturated medium chain fatty acids (i.e., greater than 50%of the fatty acids are medium chain saturated fatty acids). In someembodiments, the solubilizing agent comprises predominantly C6 to C12saturated fatty acids or derivatives of fatty acids.

In some embodiments, the solubilizing agent comprises one or more mono-,di-, or triglycerides or combinations thereof. Exemplary glycerin basedsolubilizing agents include MIGLYOLs®, which are caprylic/caprictriglycerides (SASOL Germany GMBH, Hamburg). MIGLYOLs® includes MIGLYOL®810 (caprylic/capric triglyceride), MIGLYOL® 812 (caprylic/caprictriglyceride), MIGLYOL® 816 (caprylic/capric triglyceride), and MIGLYOL®829 (caprylic/capric/succinic triglyceride). Other caprylic/caprictriglyceride solubilizing agents are likewise contemplated, including,for example: caproic/caprylic/capric/lauric triglycerides;caprylic/capric/linoleic triglycerides; or caprylic/capric/succinictriglycerides. Other exemplary caprylic/capric mono-, di-, ortriiglyceride solubilizing agents include CAPMULs® (ABITEC, Columbus,Ohio), including, but are not limited to, CAPMUL® MCM, CAPMUL® MCM C10,CAPMUL® MCM C8, CAPMUL® MCM C8 EP, and CAPMUL® 708 G. Other mono-, di-,and triglycerides of fractionated vegetable fatty acids, andcombinations or derivatives thereof can be the solubilizing agent,according to embodiments. For example, the solubilizing agent can be1,2,3-propanetriol (glycerol, glycerin, glycerine) esters of saturatedcoconut and palm kernel oil and derivatives thereof.

In some embodiments, the solubilizing agent comprises one or more estersof propylene glycol, polyethylene glycol, or combinations thereof.Exemplary propylene and polyethylene glycol based solubilizing agentsinclude glyceryl mono- and di-caprylates; propylene glycol monocaprylate(e.g., CAPMUL® PG-8 or CAPMUL® PG-8 NF); propylene glycol monocaprate(e.g., CAPMUL® PG-10); propylene glycol monolaurate (e.g., CAPMUL® PG-12EP/NF); propylene glycol mono- and dicaprylates; propylene glycol mono-and dicaprate; propylene glycol dicaprylate/dicaprate (e.g., MIGLYOL®840); propylene glycol dilaurate (e.g., CAPMUL® PG-2L EP/NF); diethyleneglycol mono ester (e.g., TRANSCUTOL®, 2-(2-Ethoxyethoxy)ethanol,GATTEFOSSÉ SAS, Saint-Priest, France); and diethylene glycol monoethylether.

In some embodiments, commercially available fatty acid glycerol andglycol ester solubilizing agents are prepared from natural oils andtherefore may comprise components in addition to the fatty acid estersthat predominantly comprise and characterize the solubilizing agent.Such other components may be, e.g., other fatty acid mono-, di-, andtriglycerides, fatty acid mono- and diester ethylene or propyleneglycols, free glycerols or glycols, or free fatty acids. For example,the Technical Data Sheet by ABITEC for CAPMUL® MCM C8 describes CAPMUL®MCM C8 as being composed of mono- and diglycerides of medium chain fattyacids (mainly caprylic) and describes the alkyl content as ≤1% C6, ≥95%C8, ≤5% C10, and ≤1.5% C12 and higher. By way of further example,MIGLYOL® 812 is generally described as a C8-C10 triglyceride because thefatty acid composition is at least about 80% caprylic (C8) acid andcapric (C10) acid. However, it can also comprise small amounts of otherfatty acids, e.g., less than about 5% of caproic (C6) acid, lauric (C12)acid, and myristic (C14) acid.

In some embodiments, the pharmaceutical composition comprises about 20%to about 85% solubilizing agent by weight, e.g., about 60% to about 85%solubilizing agent by weight. In some embodiments, the compositioncomprises progesterone, e.g., dissolved and micronized, from about 20 toabout 50 wt %, e.g., about 30 to about 35 wt %. In some embodiments, thecomposition comprises estradiol from about 0.1 to about 0.8 wt %, e.g.,about 0.15 to about 0.40 wt %.

Surfactants

In some embodiments, the pharmaceutical composition further comprisesone or more non-ionic or ionic surfactants. In some embodiments, thenon-ionic surfactant is selected from one or more of glycerol andpolyethylene glycol esters of medium chain fatty acids or long chainfatty acids, for example, lauroyl macrogol-32 glycerides or lauroylpolyoxyl-32 glycerides, commercially available as GELUCIRE®, including,for example, GELUCIRE® 39/01 (glycerol esters of saturated C12-C18 fattyacids); GELUCIRE® 43/01 (hard fat NF/JPE); GELUCIRE® 44/14 (lauroylmacrogol-32 glycerides EP, lauroyl polyoxyl-32 glycerides NF, lauroylpolyoxylglycerides (USA FDA IIG)); and GELUCIRE® 50/13 (stearoylmacrogol-32 glycerides EP, stearoyl polyoxyl-32 glycerides NF, stearoylpolyoxylglycerides (USA FDA IIG)).

In some embodiments, non-ionic surfactants comprise combinations ofmono- and di-propylene and ethylene glycols and mono-, di-, andtriglyceride combinations. For example, in some embodiments,polyethylene glycol glyceride (GELUCIRE®, GATTEFOSSE SAS, Saint-Priest,France) can be used herein as the surfactant. For example, GELUCIRE®44/14 (PEG-32 glyceryl laurate EP), a medium chain fatty acid esters ofpolyethylene glycol, is a polyethylene glycol glyceride composed ofmono-, di- and triglycerides and mono- and diesters of polyethyleneglycol.

In some embodiments, non-ionic surfactants include, for example andwithout limitation: one or more of oleic acid, linoleic acid, palmiticacid, and stearic acid. In some embodiments, non-ionic surfactantscomprise polyethylene sorbitol esters, including polysorbate 80, whichis commercially available under the trademark TWEEN 80® (Sigma Aldrich,St. Louis, Mo.). Polysorbate 80 comprises approximately 60%-70% oleicacid with the remainder comprising primarily linoleic acids, palmiticacids, and stearic acids.

In some embodiments, non-ionic surfactants include PEG-6 palmitostearateand ethylene glycol palmitostearate, which are available commercially asTEFOSE® 63 (GATTEFOSSÉ SAS, Saint-Priest, France). which can be usedwith, for example, CAPMUL® MCM having ratios of MCM to TEFOSE® 63 of,for example, 8:2 or 9:1. Other exemplary solubilizing agents/non-ionicsurfactants combinations include, without limitation: MIGLYOL®812:GELUCIRE 50/13 or MIGLYOL® 812:TEFOSE® 63.

A non-ionic or ionic surfactant may be used at concentrations greaterthan about 0.01%, for example at a concentration of about 0.01%-10.0%,about 0.1% to 10.0%, or about 1% to 10.0%. In some embodiments, thepharmaceutical composition comprises about 10.0% surfactant by weight.In some embodiments, the pharmaceutical composition comprises about 0.1%to about 5.0% surfactant by weight, e.g., about 1.0 wt %.

Other Excipients

In some embodiments, the pharmaceutical composition further comprisesone more other excipients, such as but not limited to colorants,flavoring agents, preservatives, and taste-masking agents. The choice ofexcipients will, to a large extent, depend on factors such as theparticular mode of administration, the effect of the excipients onsolubility and stability, and the nature of the dosage form. Colorants,for example, may comprise about 0.1% to about 2% by weight.Preservatives may comprise methyl and propyl paraben, for example, in aratio of about 10:1, and at a proportion of about 0.005% and 0.05% byweight.

Generally, the solubilizing agents, surfactants, and excipients used inthe pharmaceutical compositions described herein are non-toxic,pharmaceutically acceptable, compatible with each other, and maintainstability of the pharmaceutical composition and the various componentswith respect to each other. Additionally, the combination of variouscomponents that comprise the pharmaceutical compositions will maintainwill result in the desired therapeutic effect when administered to asubject.

Formulation

In some embodiments, combinations of solubilizing agents (e.g., two ormore oils) or combinations of one or more solubilizing agents and one ormore surfactants are used to form estradiol and progesteronecompositions. Various ratios of these solubilizing agents orsolubilizing agents and surfactants can be used. For example, CAPMUL®MCM and a non-ionic surfactant, e.g., GELUCIRE® 44/14 (lauroylmacrogol-32 glycerides EP; lauroyl polyoxyl-32 glycerides NF; lauroylpolyoxylglycerides (USA FDA IIG)), can be used at ratios of about 99:1to about 2:1, including, for example and without limitation: 60:40,65:35, 70:30, 75:25, 80:10, 80:15, 85:20, 90:10, and 98:1. As anotherexample, CAPMUL® MCM and a non-ionic surfactant, e.g., TEFOSE® 63, canbe used as rations of about 8:2 or 9:1. Other exemplary solubilizingagent/surfactant combinations include, without limitation: MIGLYOL®812:GELUCIRE® 50/13 or MIGLYOL® 812:TEFOSE® 63. The ratios of oil (e.g.,medium chain fatty acid esters of monoglycerides and diglycerides) tonon-ionic surfactant can be significantly higher. For example, CAPMUL®MCM and GELUCIRE® can be used in ratios of up to about 65:1, e.g., 8:1,22:1, 49:1, 65:1 and 66:1. Thus, useful ratios can be 8:1 or greater,e.g., 60 to 70:1.

In some embodiments, estradiol or progesterone is soluble in thesolubilizing agent at room temperature, although it may be desirable towarm certain solubilizing agents. For example, when the formulationcomprises medium chain fatty acid mono- and diglycerides (e.g., CAPMUL®MCM) and polyethylene glycol glycerides (e.g., GELUCIRE®) as asurfactant, the oil or the surfactant can be warmed up, e.g., to about65° C. for the surfactant and less for the oil, to facilitate mixing ofthe oil and surfactant. The estradiol can be added at this temperature,or at lower temperatures as the mixture cools, e.g., about 40° C. orabout 30° C., or even after the mixture has cooled to room temperature.The progesterone can also be added as the mixture cools, e.g., to belowabout 40° C. or to below about 30° C., or after the mixture has cooledto room temperature.

As a non-limiting example, a composition of this disclosure comprisessolubilized estradiol; progesterone, at least 30% (e.g., at least about30%, about 40%, about 50%, about 60%, about 70%, about 75%, about 80%,about 85%, or more) of the progesterone being solubilized (the balancebeing micronized as discussed elsewhere herein); and a solubilizingagent that is an oil, wherein the oil comprises medium chain fatty acidmono-, di-, or triglycerides, with or without a surfactant. In certainembodiments, a specification for progesterone is set at >80%solubilized, <20% micronized or >85% solubilized, <15% micronized.Specific examples of such illustrative embodiments, with CAPMUL® MCM NF(glyceryl caprylate/caprate) as a solubilizing agent and GELUCIRE® 44/14(lauroyl polyoxyglyceride) as a surfactant, in which at least about 85%of the progesterone can be solubilized, include, e.g., the followingfive formulations A-E:

TABLE 1 Pharmaceutical Composition A - progesterone 50 mg/estradiol 0.25mg Ingredient Amount (% w/w) Qty/Capsule (mg) Progesterone, USP, 33.3350.00 micronized Estradiol Hemihydrate 0.17 0.26 CAPMUL ® MCM, NF 65.4998.24 GELUCIRE ® 44/14, NF 1.00 1.50 Total 100.00 150.00

TABLE 2 Pharmaceutical Composition B - progesterone 50 mg/estradiol 0.5mg Ingredient Amount (% w/w) Qty/Capsule (mg) Progesterone, USP, 33.3350.00 micronized Estradiol Hemihydrate 0.35 0.52 CAPMUL ® MCM, NF 65.3297.98 GELUCIRE ® 44/14, NF 1.00 1.50 Total 100.00 150.00

TABLE 3 Pharmaceutical Composition C - progesterone 100 mg/estradiol 0.5mg Ingredient Amount (% w/w) Qty/Capsule (mg) Progesterone, USP, 33.33100.00 micronized Estradiol Hemihydrate 0.17 0.52 CAPMUL ® MCM, NF 65.49196.48 GELUCIRE ® 44/14, NF 1.00 3.00 Total 100.00 300.00

TABLE 4 Pharmaceutical Composition D - progesterone 100 mg/estradiol 1mg Ingredient Amount (% w/w) Qty/Capsule (mg) Progesterone, USP, 33.33100.00 micronized Estradiol Hemihydrate 0.34 1.03 CAPMUL ® MCM, NF 65.32195.97 GELUCIRE ® 44/14, NF 1.00 3.00 Total 100.00 300.00

TABLE 5 Pharmaceutical Composition E - progesterone 200 mg/estradiol 2mg Ingredient Amount (% w/w) Qty/Capsule (mg) Progesterone, USP, 33.33200.00 micronized Estradiol Hemihydrate 0.34 2.06 CAPMUL ® MCM, NF 65.32391.94 GELUCIRE ® 44/14, NF 1.00 6.00 Total 100.00 600.00 * Note: 1.00mg Estradiol is equivalent to 1.03 mg Estradiol Hemihydrate

In general terms, the above formulations comprise 30 to 35 wt %progesterone, 0.1 to 0.4 wt % estradiol (or estradiol hemihydrate), 55to 75 wt % of an oil that is predominantly medium chain fatty acidmono-, di-, or triglycerides, such as CAPMUL® MCM, and 0.5 to 10 wt % ofa non-ionic surfactant, such as GELUCIRE® 44/14. The above formulationsmay be modified to comprise excipients, e.g., gelatin such as Gelatin200 Bloom, glycerin, coloring agents such as Opatint red and white, and,optionally, MIGLYOL® 812.

Estradiol solubilization helps ensure high content uniformity andenhanced stability. Fully solubilized progesterone formulations orpartially solubilized progesterone formulations in which at least about50% of the progesterone, e.g., at least about 50%, 60%, 70%, 75%, 80%,85%, 90%, 95% or more, is solubilized appear to provide improvedPK-related properties.

Pharmacokinetic Parameters of Estradiol and Progesterone Compositions

The pharmaceutical compositions of this disclosure can be formulated toprovide desirable pharmacokinetic parameters in a subject (e.g., afemale subject) to whom the composition is administered. In someembodiments, a pharmaceutical composition as described herein producesdesirable pharmacokinetic parameters for progesterone in the subject. Insome embodiments, a pharmaceutical composition as described hereinproduces desirable pharmacokinetic parameters for estradiol in thesubject. In some embodiments, a pharmaceutical composition as describedherein produces desirable pharmacokinetic parameters for one or moremetabolites of progesterone or estradiol in the subject, for example,estrone or total estrone.

Following the administration of a composition comprising progesteroneand estradiol to a subject, the concentration and metabolism ofprogesterone or estradiol can be measured in a sample (e.g., a blood,serum, or plasma sample) from the subject. Progesterone is metabolizedto pregnanediols and pregnanolones, which are then conjugated toglucuronide and sulfate metabolites that are excreted or furtherrecycled. Estradiol is converted reversibly to estrone, and bothestradiol and estrone can be converted to the metabolite estriol. Inpostmenopausal women, a significant proportion of circulating estrogensexist as sulfate conjugates, especially estrone sulfate. Thus, estronecan be measured with respect to “estrone” amounts (excluding conjugatessuch as estrone sulfate) and “total estrone” amounts (including bothfree, or unconjugated, estrone and conjugated estrone such as estronesulfate).

The pharmaceutical compositions of this disclosure can be characterizedfor one or more pharmacokinetic parameters of progesterone, estradiol,or a metabolite thereof following administration of the composition to asubject or to a population of subjects. These pharmacokinetic parametersinclude AUC, C_(max), and T_(max). AUC is a determination of the areaunder the curve (AUC) plotting the blood, serum, or plasma concentrationof drug along the ordinate (Y-axis) against time along the abscissa(X-axis). AUCs are well understood, frequently used tools in thepharmaceutical arts and have been extensively described. C_(max) is wellunderstood in the art as an abbreviation for the maximum drugconcentration in blood, serum, or plasma of a subject. T_(max) is wellunderstood in the art as an abbreviation for the time to maximum drugconcentration in blood, serum, or plasma of a subject.

In some embodiments, one or more pharmacokinetic parameters, e.g., AUC,C_(max), or T_(max), is measured for estradiol. In some embodiments, oneor more pharmacokinetic parameters, e.g., AUC, C_(max), or T_(max), ismeasured for progesterone. In some embodiments, one or morepharmacokinetic parameters, e.g., AUC, C_(max), or T_(max), is measuredfor estrone. In some embodiments, one or more pharmacokineticparameters, e.g., AUC, C_(max), or T_(max), is measured for totalestrone.

Any of a variety of methods can be used for measuring the levels ofprogesterone, estradiol, estrone, or total estrone in a sample,including immunoassays, mass spectrometry (MS), high performance liquidchromatography (HPLC) with ultraviolet fluorescent detection, liquidchromatography in conjunction with mass spectrometry (LC-MS), tandemmass spectrometry (MS/MS), and liquid chromatography-tandem massspectrometry (LC-MS/MS). In some embodiments, the levels ofprogesterone, estradiol, estrone, or total estrone are measured using avalidated LC-MS/MS method. Methods of measuring hormone levels are welldescribed in the literature.

The levels of progesterone, estradiol, estrone, or total estrone can bemeasured in any biological sample, e.g. a tissue or fluid such as blood,serum, plasma, or urine. In some embodiments, the sample is blood orplasma. In some embodiments, the levels of progesterone, estradiol,estrone, or total estrone are measured about 0.0, 0.10, 0.20, 0.05,0.30, 0.35, 0.40, 0.45, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 15, 18,21, 24, 27, 30, 33, 36, 39, 42, 45, or 48 hours after dosing, or anyother appropriate time period that is common or useful in determiningthe levels of each of the hormones. In some embodiments, the levels ofprogesterone, estradiol, estrone, or total estrone are measured about 18hours, about 24 hours, about 18-36 hours, about 20-30 hours, about 22-26hours, about 24-36 hours, about 36 hours, about 36-48 hours, about 40-48hours, or about 48 hours after administration of a single dose or afirst dose. Generally, assays to determine the levels of progesterone,estradiol, estrone, or total estrone are measured one or more timesevery 5, 10, 15, 20, 30, 60, 120, 360, 480, 720, or 1440 minutes afteradministration, or combinations thereof (e.g., the first measurementsare taken every 15 minutes for the first hour, followed by every 120minutes thereafter). In embodiments, the timing of such measurements aredesigned to accurately measure C_(max), T_(max), or AUC. Timing can beadjusted based on the given circumstances (i.e., one formulation maycause a more rapid C_(max), in which case the initial times would beclustered closer together, closer to time zero, or both to ensureaccurate measurement of C_(max), T_(max), and AUC). In some embodiments,the C_(max), T_(max), or AUC values for progesterone, estradiol,estrone, or total estrone are measured following administration of asingle dose of a pharmaceutical composition as described herein.

In some embodiments, the values for C_(max), T_(max), or AUC represent anumber of values taken from all the subjects in a patient population andare, therefore, mean values (e.g., arithmetic or geometric means)averaged over the entire population.

In some embodiments, oral administration of a pharmaceutical compositioncomprising estradiol, progesterone, and a medium chain solubilizingagent as described herein to a subject, or to a population of subjects,produces one or more AUC, C_(max), or T_(max) parameters, or one or moremean AUC, mean C_(max), or mean T_(max) parameters, respectively, forestradiol, progesterone, estrone, or total estrone as described below.

AUC, C_(max), and T_(max) parameters (A)

In some embodiments, a pharmaceutical composition of this disclosurecomprises estradiol at a dosage of about 0.25 mg and progesterone at adosage of about 50 mg. In some embodiments, the pharmaceuticalcomposition comprises the formulation of Formulation A in Table 1 above.

In some embodiments, administration of a composition comprising about0.25 mg estradiol and about 50 mg progesterone to a subject produces, ina plasma sample from the subject, one or both parameters selected from:

-   -   (i) an AUC_((0-t)) for estradiol that is from 140.3733 pg·hr/ml        to 219.3333 pg·hr/ml; or    -   (ii) a C_(max) for estradiol that is from 6.4790 pg/ml to        10.1235 pg/ml.

In some embodiments, administration of the composition to the subjectproduces both an AUC_((0-t)) for estradiol that is from 140.3733pg·hr/ml to 219.3333 pg·hr/ml, and a C_(max) for estradiol that is from6.4790 pg/ml to 10.1235 pg/ml.

In some embodiments, administration of the composition to the subjectfurther produces, in a plasma sample from the subject, one or bothparameters selected from:

-   -   (i) an AUC_((0-t)) for progesterone that is from 24.0174        ng·hr/ml to 37.5272 ng·hr/ml; or    -   (ii) a C_(max) for progesterone that is from 17.8444 ng/ml to        27.8819 ng/ml.

In some embodiments, administration of the composition to the subjectproduces both an AUC_((0-t)) for progesterone that is from 24.0174ng·hr/ml to 37.5272 ng·hr/ml, and a C_(max) for progesterone that isfrom 17.8444 ng/ml to 27.8819 ng/ml.

In some embodiments, administration of the composition to the subjectproduces, in a plasma sample from the subject,

-   -   (i) an AUC_((0-t)) for estradiol that is from 140.3733 pg·hr/ml        to 219.3333 pg·hr/ml;    -   (ii) a C_(max) for estradiol that is from 6.4790 pg/ml to        10.1235 pg/ml;    -   (iii) an AUC_((0-t)) for progesterone that is from 24.0174        ng·hr/ml to 37.5272 ng·hr/ml; or    -   (iv) a C_(max) for progesterone that is from 17.8444 ng/ml to        27.8819 ng/ml.

In some embodiments, administration of the composition to the subjectfurther produces, in a plasma sample from the subject, a T_(max) forestradiol that is from 7.2 hr to 11.3 hr. In some embodiments,administration of the composition to the subject further produces, in aplasma sample from the subject, a T_(max) for progesterone that is from2.4 hr to 3.8 hr.

In some embodiments, administration of the pharmaceutical composition tothe subject produces, in a plasma sample from the subject, one, two,three or more parameters selected from:

-   -   (i) an AUC_((0-t)) for estradiol that is from 140.3733 pg·hr/ml        to 219.3333 pg·hr/ml;    -   (ii) a C_(max) for estradiol that is from 6.4790 pg/ml to        10.1235 pg/ml;    -   (iii) an AUC_((0-t)) for progesterone that is from 24.0174        ng·hr/ml to 37.5272 ng·hr/ml; or    -   (iv) a C_(max) for progesterone that is from 17.8444 ng/ml to        27.8819 ng/ml.

In some embodiments, administration of the pharmaceutical composition tothe subject produces both parameters (i) and (ii). In some embodiments,administration of the composition to the subject produces bothparameters (i) and (iii). In some embodiments, administration of thecomposition to the subject produces both parameters (i) and (iv). Insome embodiments, administration of the composition to the subjectproduces both parameters (ii) and (iii). In some embodiments,administration of the composition to the subject produces bothparameters (ii) and (iv). In some embodiments, administration of thecomposition to the subject produces both parameters (iii) and (iv). Insome embodiments, administration of the composition to the subjectproduces all of parameters (i), (ii), and (iii). In some embodiments,administration of the composition to the subject produces bothparameters (i), (iii), and (iv). In some embodiments, administration ofthe composition to the subject produces both parameters (ii), (iii), and(iv). In some embodiments, administration of the composition to thesubject produces all of parameters (i), (ii), (iii), and (iv).

In some embodiments, administration of the pharmaceutical composition tothe subject further produces, in a plasma sample from the subject, oneor more parameters selected from:

-   -   (i) an AUC_((0-t)) for estrone that is from 909.6091 pg·hr/ml to        1421.2642 pg·hr/ml;    -   (ii) a C_(max) for estrone that is from 42.6549 pg/ml to 66.6483        pg/ml; or    -   (iii) a T_(max) for estrone that is from 4.4 hr to 6.9 hr.

In some embodiments, administration of the pharmaceutical composition tothe subject further produces, in a plasma sample from the subject, oneor more parameters selected from:

-   -   (i) an AUC_((0-t)) for total estrone that is from 20.1752        ng·hr/ml to 31.5238 ng·hr/ml;    -   (ii) a C_(max) for total estrone that is from 3.5429 ng/ml to        5.5358 ng/ml; or    -   (iii) a T_(max) for total estrone that is from 2 hr to 3.1 hr.

In some embodiments, a pharmaceutical composition comprising about 0.25mg estradiol and about 50 mg progesterone is administered to apopulation of subjects in need thereof, and mean parameters aredetermined for samples (e.g., blood or plasma samples) from the subjectsadministered the composition. Thus, in some embodiments, administrationof the composition to a population of subject produces, in plasmasamples from the subjects, one or more of a mean AUC_((0-t)) forestradiol that is from 140.3733 pg·hr/ml to 219.3333 pg·hr/ml, a meanC_(max) for estradiol that is from 6.4790 pg/ml to 10.1235 pg/ml, and amean T_(max) for estradiol that is from 7.2 hr to 11.3 hr. In someembodiments, administration of the composition to a population ofsubject produces, in plasma samples from the subjects, one or more of amean AUC_((0-t)) for progesterone that is from 24.0174 ng·hr/ml to37.5272 ng·hr/ml, a mean C_(max) for progesterone that is from 17.8444ng/ml to 27.8819 ng/ml, and a mean T_(max) for progesterone that is from2.4 hr to 3.8 hr. In some embodiments, administration of the compositionto a population of subject produces, in plasma samples from thesubjects, one or more of a mean AUC_((0-t)) for estrone that is from909.6091 pg·hr/ml to 1421.2642 pg·hr/ml, a mean C_(max) for estrone thatis from 42.6549 pg/ml to 66.6483 pg/ml, and a mean T_(max) for estronethat is from 4.4 hr to 6.9 hr.

In some embodiments, administration of the composition to a populationof subject produces, in plasma samples from the subjects, one or more ofa mean AUC_((0-t)) for total estrone that is from 20.1752 ng·hr/ml to31.5238 ng·hr/ml, a mean C_(max) for total estrone that is from 3.5429ng/ml to 5.5358 ng/ml, and a mean T_(max) for total estrone that is from2 hr to 3.1 hr.

In some embodiments, methods of treating a subject with a pharmaceuticalcomposition comprising estradiol and progesterone are provided. In someembodiments, the method comprises administering to the subject apharmaceutical composition comprising about 0.25 mg estradiol and about50 mg progesterone as described herein (e.g., a pharmaceuticalcomposition having the formulation of Formulation A in Table 1 above),wherein administration of the pharmaceutical composition produces, in aplasma sample from the subject, one or more parameters selected from: anAUC_((0-t)) for estradiol that is from 140.3733 pg·hr/ml to 219.3333pg·hr/ml; a C_(max) for estradiol that is from 6.4790 pg/ml to 10.1235pg/ml; a T_(max) for estradiol that is from 7.2 hr to 11.3 hr; anAUC_((0-t)) for progesterone that is from 24.0174 ng·hr/ml to 37.5272ng·hr/ml; a C_(max) for progesterone that is from 17.8444 ng/ml to27.8819 ng/ml; a T_(max) for progesterone that is from 2.4 hr to 3.8 hr;an AUC_((0-t)) for estrone that is from 909.6091 pg·hr/ml to 1421.2642pg·hr/ml; a C_(max) for estrone that is from 42.6549 pg/ml to 66.6483pg/ml; a T_(max) for estrone that is from 4.4 hr to 6.9 hr; anAUC_((0-t)) for total estrone that is from 20.1752 ng·hr/ml to 31.5238ng·hr/ml; a C_(max) for total estrone that is from 3.5429 ng/ml to5.5358 ng/ml; and a T_(max) for total estrone that is from 2 hr to 3.1hr.

In some embodiments, the method further comprises obtaining a samplefrom the subject (e.g., a blood or plasma sample) followingadministration of a single dose of the pharmaceutical composition (e.g.,a pharmaceutical composition having the formulation of Formulation A inTable 1 above), and measuring one or more pharmacokinetic parametersselected from an AUC_((0-t)) for estradiol, a C_(max) for estradiol, anAUC_((0-t)) for progesterone, a C_(max) for progesterone, an AUC_((0-t))for estrone, a C_(max) for estrone, an AUC_((0-t)) for total estrone,and a C_(max) for total estrone; wherein the presence of one or more ofthe following values is indicative of a therapeutically effective dose:an AUC_((0-t)) for estradiol that is from 140.3733 pg·hr/ml to 219.3333pg·hr/ml; a C_(max) for estradiol that is from 6.4790 pg/ml to 10.1235pg/ml; an AUC_((0-t)) for progesterone that is from 24.0174 ng·hr/ml to37.5272 ng·hr/ml; a C_(max) for progesterone that is from 17.8444 ng/mlto 27.8819 ng/ml; an AUC_((0-t)) for estrone that is from 909.6091pg·hr/ml to 1421.2642 pg·hr/ml; a C_(max) for estrone that is from42.6549 pg/ml to 66.6483 pg/ml; an AUC_((0-t)) for total estrone that isfrom 20.1752 ng·hr/ml to 31.5238 ng·hr/ml; or a C_(max) for totalestrone that is from 3.5429 ng/ml to 5.5358 ng/ml. In some embodiments,the one or more pharmacokinetic parameters are measured about 18 hours,about 24 hours, about 18-36 hours, about 20-30 hours, about 22-26 hours,about 24-36 hours, about 36 hours, about 36-48 hours, about 40-48 hours,or about 48 hours after administration of the single dose.

AUC, C_(max), and T_(max) Parameters (B)

In some embodiments, a pharmaceutical composition of this disclosurecomprises estradiol at a dosage of about 0.50 mg and progesterone at adosage of about 50 mg. In some embodiments, the pharmaceuticalcomposition comprises the formulation of Formulation B in Table 2 above.

In some embodiments, administration of a composition comprising about0.50 mg estradiol and about 50 mg progesterone to a subject produces, ina plasma sample from the subject, one or both parameters selected from:

-   -   (i) an AUC_((0-t)) for estradiol that is from 280.7467 pg·hr/ml        to 438.6667 pg·hr/ml; or    -   (ii) a C_(max) for estradiol that is from 12.9580 pg/ml to        20.2469 pg/ml.

In some embodiments, administration of the composition to the subjectproduces both an AUC_((0-t)) for estradiol that is from 280.7467pg·hr/ml to 438.6667 pg·hr/ml, and a C_(max) for estradiol that is from12.9580 pg/ml to 20.2469 pg/ml.

In some embodiments, administration of the composition to the subjectfurther produces, in a plasma sample from the subject, one or bothparameters selected from:

-   -   (i) an AUC_((0-t)) for progesterone that is from 24.0174        ng·hr/ml to 37.5272 ng·hr/ml; or    -   (ii) a C_(max) for progesterone that is from 17.8444 ng/ml to        27.8819 ng/ml.

In some embodiments, administration of the composition to the subjectproduces both an (AUC)_((0-t)) for progesterone that is from 24.0174ng·hr/ml to 37.5272 ng·hr/ml, and a C_(max) for progesterone that isfrom 17.8444 ng/ml to 27.8819 ng/ml.

In some embodiments, administration of the composition to the subjectproduces, in a plasma sample from the subject,

-   -   (i) an AUC_((0-t)) for estradiol that is from 280.7467 pg·hr/ml        to 438.6667 pg·hr/ml;    -   (ii) a C_(max) for estradiol that is from 12.9580 pg/ml to        20.2469 pg/ml;    -   (iii) an AUC_((0-t)) for progesterone that is from 24.0174        ng·hr/ml to 37.5272 ng·hr/ml; or    -   (iv) a C_(max) for progesterone that is from 17.8444 ng/ml to        27.8819 ng/ml.

In some embodiments, administration of the composition to the subjectfurther produces, in a plasma sample from the subject, a T_(max) forestradiol that is from 7.2 hr to 11.3 hr. In some embodiments,administration of the composition to the subject further produces, in aplasma sample from the subject, a T_(max) for progesterone that is from2.4 hr to 3.8 hr.

In some embodiments, administration of the pharmaceutical composition tothe subject produces, in a plasma sample from the subject, one, two,three or more parameters selected from:

-   -   (i) an AUC_((0-t)) for estradiol that is from 280.7467 pg·hr/ml        to 438.6667 pg·hr/ml;    -   (ii) a C_(max) for estradiol that is from 12.9580 pg/ml to        20.2469 pg/ml;    -   (iii) an AUC_((0-t)) for progesterone that is from 24.0174        ng·hr/ml to 37.5272 ng·hr/ml; or    -   (iv) a C_(max) for progesterone that is from 17.8444 ng/ml to        27.8819 ng/ml.

In some embodiments, administration of the pharmaceutical composition tothe subject produces both parameters (i) and (ii). In some embodiments,administration of the composition to the subject produces bothparameters (i) and (iii). In some embodiments, administration of thecomposition to the subject produces both parameters (i) and (iv). Insome embodiments, administration of the composition to the subjectproduces both parameters (ii) and (iii). In some embodiments,administration of the composition to the subject produces bothparameters (ii) and (iv). In some embodiments, administration of thecomposition to the subject produces both parameters (iii) and (iv). Insome embodiments, administration of the composition to the subjectproduces all of parameters (i), (ii), and (iii). In some embodiments,administration of the composition to the subject produces bothparameters (i), (iii), and (iv). In some embodiments, administration ofthe composition to the subject produces both parameters (ii), (iii), and(iv). In some embodiments, administration of the composition to thesubject produces all of parameters (i), (ii), (iii), and (iv).

In some embodiments, administration of the pharmaceutical composition tothe subject further produces, in a plasma sample from the subject, oneor more parameters selected from:

-   -   (i) an AUC_((0-t)) for estrone that is from 1819.2181 pg·hr/ml        to 2842.5283 pg·hr/ml;    -   (ii) a C_(max) for estrone that is from 85.3098 pg/ml to        133.2966 pg/ml; or    -   (iii) a T_(max) for estrone that is from 4.4 hr to 6.9 hr.

In some embodiments, administration of the pharmaceutical composition tothe subject further produces, in a plasma sample from the subject, oneor more parameters selected from:

-   -   (i) an AUC_((0-t)) for total estrone that is from 40.3505        ng·hr/ml to 63.0476 ng·hr/ml;    -   (ii) a C_(max) for total estrone that is from 7.0858 ng/ml to        11.0715 ng/ml; or    -   (iii) a T_(max) for total estrone that is from 2 hr to 3.1 hr.

In some embodiments, a pharmaceutical composition comprising about 0.50mg estradiol and about 50 mg progesterone is administered to apopulation of subjects in need thereof, and mean parameters aredetermined for samples (e.g., blood or plasma samples) from the subjectsadministered the composition. Thus, in some embodiments, administrationof the composition to a population of subject produces, in plasmasamples from the subjects, one or more of a mean AUC_((0-t)) forestradiol that is from 280.7467 pg·hr/ml to 438.6667 pg·hr/ml, a meanC_(max) for estradiol that is from 12.9580 pg/ml to 20.2469 pg/ml, and amean T_(max) for estradiol that is from 7.2 hr to 11.3 hr. In someembodiments, administration of the composition to a population ofsubject produces, in plasma samples from the subjects, one or more of amean AUC_((0-t)) for progesterone that is from 24.0174 ng·hr/ml to37.5272 ng·hr/ml, a mean C_(max) for progesterone that is from 17.8444ng/ml to 27.8819 ng/ml, and a mean T_(max) for progesterone that is from2.4 hr to 3.8 hr. In some embodiments, administration of the compositionto a population of subject produces, in plasma samples from thesubjects, one or more of a mean AUC_((0-t)) for estrone that is from1819.2181 pg·hr/ml to 2842.5283 pg·hr/ml, a mean C_(max) for estronethat is from 85.3098 pg/ml to 133.2966 pg/ml, and a mean T_(max) forestrone that is from 4.4 hr to 6.9 hr. In some embodiments,administration of the composition to a population of subject produces,in plasma samples from the subjects, one or more of a mean AUC_((0-t))for total estrone that is from 40.3505 ng·hr/ml to 63.0476 ng·hr/ml, amean C_(max) for total estrone that is from 7.0858 ng/ml to 11.0715ng/ml, and a mean T_(max) for total estrone that is from 2 hr to 3.1 hr.

In some embodiments, methods of treating a subject with a pharmaceuticalcomposition comprising estradiol and progesterone are provided. In someembodiments, the method comprises administering to the subject apharmaceutical composition comprising about 0.50 mg estradiol and about50 mg progesterone as described herein (e.g., a pharmaceuticalcomposition having the formulation of Formulation B in Table 2 above),wherein administration of the pharmaceutical composition produces, in aplasma sample from the subject, one or more parameters selected from: anAUC_((0-t)) for estradiol that is from 280.7467 pg·hr/ml to 438.6667pg·hr/ml; a C_(max) for estradiol that is from 12.9580 pg/ml to 20.2469pg/ml; a T_(max) for estradiol that is from 7.2 hr to 11.3 hr; anAUC_((0-t)) for progesterone that is from 24.0174 ng·hr/ml to 37.5272ng·hr/ml; a C_(max) for progesterone that is from 17.8444 ng/ml to27.8819 ng/ml; a T_(max) for progesterone that is from 2.4 hr to 3.8 hr;an AUC_((0-t)) for estrone that is from 1819.2181 pg·hr/ml to 2842.5283pg·hr/ml; a C_(max) for estrone that is from 85.3098 pg/ml to 133.2966pg/ml; a T_(max) for estrone that is from 4.4 hr to 6.9 hr; anAUC_((0-t)) for total estrone that is from 40.3505 ng·hr/ml to 63.0476ng·hr/ml; a C_(max) for total estrone that is from 7.0858 ng/ml to11.0715 ng/ml; and a T_(max) for total estrone that is from 2 hr to 3.1hr.

In some embodiments, the method further comprises obtaining a samplefrom the subject (e.g., a blood or plasma sample) followingadministration of a single dose of the pharmaceutical composition (e.g.,a pharmaceutical composition having the formulation of Formulation B inTable 2 above), and measuring one or more pharmacokinetic parametersselected from an AUC_((0-t)) for estradiol, a C_(max) for estradiol, anAUC_((0-t)) for progesterone, a C_(max) for progesterone, an AUC_((0-t))for estrone, a C_(max) for estrone, an AUC_((0-t)) for total estrone,and a C_(max) for total estrone; wherein the presence of one or more ofthe following values is indicative of a therapeutically effective dose:an AUC_((0-t)) for estradiol that is from 280.7467 pg·hr/ml to 438.6667pg·hr/ml; a C_(max) for estradiol that is from 12.9580 pg/ml to 20.2469pg/ml; an AUC_((0-t)) for progesterone that is from 24.0174 ng·hr/ml to37.5272 ng·hr/ml; a C_(max) for progesterone that is from 17.8444 ng/mlto 27.8819 ng/ml; an AUC_((0-t)) for estrone that is from 1819.2181pg·hr/ml to 2842.5283 pg·hr/ml; a C_(max) for estrone that is from85.3098 pg/ml to 133.2966 pg/ml; an AUC_((0-t)) for total estrone thatis from 40.3505 ng·hr/ml to 63.0476 ng·hr/ml; and a C_(max) for totalestrone that is from 7.0858 ng/ml to 11.0715 ng/ml. In some embodiments,the one or more pharmacokinetic parameters are measured about 18 hours,about 24 hours, about 18-36 hours, about 20-30 hours, about 22-26 hours,about 24-36 hours, about 36 hours, about 36-48 hours, about 40-48 hours,or about 48 hours after administration of the single dose.

AUC, C_(max), and T_(max) Parameters (C)

In some embodiments, a pharmaceutical composition of this disclosurecomprises estradiol at a dosage of about 0.50 mg and progesterone at adosage of about 100 mg. In some embodiments, the pharmaceuticalcomposition comprises the formulation of Formulation C in Table 3 above.

In some embodiments, administration of a composition comprising about0.50 mg estradiol and about 100 mg progesterone to a subject produces,in a plasma sample from the subject, one or both parameters selectedfrom:

-   -   (i) an AUC_((0-t)) for estradiol that is from 280.7467 pg·hr/ml        to 438.6667 pg·hr/ml; or    -   (ii) a C_(max) for estradiol that is from 12.9580 pg/ml to        20.2469 pg/ml.

In some embodiments, administration of the composition to the subjectproduces both an AUC_((0-t)) for estradiol that is from 280.7467pg·hr/ml to 438.6667 pg·hr/ml, and a C_(max) for estradiol that is from12.9580 pg/ml to 20.2469 pg/ml.

In some embodiments, administration of the composition to the subjectfurther produces, in a plasma sample from the subject, one or bothparameters selected from:

-   -   (i) an AUC_((0-t)) for progesterone that is from 48.0348        ng·hr/ml to 75.0543 ng·hr/ml; or    -   (ii) a C_(max) for progesterone that is from 35.6889 ng/ml to        55.7639 ng/ml.

In some embodiments, administration of the composition to the subjectproduces both an AUC_((0-t)) for progesterone that is from 48.0348ng·hr/ml to 75.0543 ng·hr/ml, and a C_(max) for progesterone that isfrom 35.6889 ng/ml to 55.7639 ng/ml.

In some embodiments, administration of the composition to the subjectproduces, in a plasma sample from the subject,

-   -   (i) an AUC_((0-t)) for estradiol that is from 280.7467 pg·hr/ml        to 438.6667 pg·hr/ml;    -   (ii) a C_(max) for estradiol that is from 12.9580 pg/ml to        20.2469 pg/ml;    -   (iii) an AUC_((0-t)) for progesterone that is from 48.0348        ng·hr/ml to 75.0543 ng·hr/ml; or    -   (iv) a C_(max) for progesterone that is from 35.6889 ng/ml to        55.7639 ng/ml.

In some embodiments, administration of the composition to the subjectfurther produces, in a plasma sample from the subject, a T_(max) forestradiol that is from 7.2 hr to 11.3 hr. In some embodiments,administration of the composition to the subject further produces, in aplasma sample from the subject, a T_(max) for progesterone that is from2.4 hr to 3.8 hr.

In some embodiments, administration of the pharmaceutical composition tothe subject produces, in a plasma sample from the subject, one or moreparameters selected from:

-   -   (i) an AUC_((0-t)) for estradiol that is from 280.7467 pg·hr/ml        to 438.6667 pg·hr/ml;    -   (ii) a C_(max) for estradiol that is from 12.9580 pg/ml to        20.2469 pg/ml;    -   (iii) an AUC_((0-t)) for progesterone that is from 48.0348        ng·hr/ml to 75.0543 ng·hr/ml; or    -   (iv) a C_(max) for progesterone that is from 35.6889 ng/ml to        55.7639 ng/ml.

In some embodiments, administration of the pharmaceutical composition tothe subject produces both parameters (i) and (ii). In some embodiments,administration of the composition to the subject produces bothparameters (i) and (iii). In some embodiments, administration of thecomposition to the subject produces both parameters (i) and (iv). Insome embodiments, administration of the composition to the subjectproduces both parameters (ii) and (iii). In some embodiments,administration of the composition to the subject produces bothparameters (ii) and (iv). In some embodiments, administration of thecomposition to the subject produces both parameters (iii) and (iv). Insome embodiments, administration of the composition to the subjectproduces all of parameters (i), (ii), and (iii). In some embodiments,administration of the composition to the subject produces bothparameters (i), (iii), and (iv). In some embodiments, administration ofthe composition to the subject produces both parameters (ii), (iii), and(iv). In some embodiments, administration of the composition to thesubject produces all of parameters (i), (ii), (iii), and (iv).

In some embodiments, administration of the pharmaceutical composition tothe subject further produces, in a plasma sample from the subject, one,two, three or more parameters selected from:

-   -   (i) an AUC_((0-t)) for estrone that is from 1819.2181 pg·hr/ml        to 2842.5283 pg·hr/ml;    -   (ii) a C_(max) for estrone that is from 85.3098 pg/ml to        133.2966 pg/ml; or    -   (iii) a T_(max) for estrone that is from 4.4 hr to 6.9 hr.

In some embodiments, administration of the pharmaceutical composition tothe subject further produces, in a plasma sample from the subject, oneor more parameters selected from:

-   -   (i) an AUC_((0-t)) for total estrone that is from 40.3505        ng·hr/ml to 63.0476 ng·hr/ml;    -   (ii) a C_(max) for total estrone that is from 7.0858 ng/ml to        11.0715 ng/ml; or    -   (iii) a T_(max) for total estrone that is from 2 hr to 3.1 hr.

In some embodiments, a pharmaceutical composition comprising about 0.50mg estradiol and about 100 mg progesterone is administered to apopulation of subjects in need thereof, and mean parameters aredetermined for samples (e.g., blood and plasma samples) from thesubjects administered the composition. Thus, in some embodiments,administration of the composition to a population of subject produces,in plasma samples from the subjects, one or more of a mean AUC_((0-t))for estradiol that is from 280.7467 pg·hr/ml to 438.6667 pg·hr/ml, amean C_(max) for estradiol that is from 12.9580 pg/ml to 20.2469 pg/ml,and a mean T_(max) for estradiol that is from 7.2 hr to 11.3 hr. In someembodiments, administration of the composition to a population ofsubject produces, in plasma samples from the subjects, one or more of amean AUC_((0-t)) for progesterone that is from 48.0348 ng·hr/ml to75.0543 ng·hr/ml, a mean C_(max) for progesterone that is from 35.6889ng/ml to 55.7639 ng/ml, and a mean T_(max) for progesterone that is from2.4 hr to 3.8 hr. In some embodiments, administration of the compositionto a population of subject produces, in plasma samples from thesubjects, one or more of a mean AUC_((0-t)) for estrone that is from1819.2181 pg·hr/ml to 2842.5283 pg·hr/ml, a mean C_(max) for estronethat is from 85.3098 pg/ml to 133.2966 pg/ml, and a mean T_(max) forestrone that is from 4.4 hr to 6.9 hr. In some embodiments,administration of the composition to a population of subject produces,in plasma samples from the subjects, one or more of a mean AUC_((0-t))for total estrone that is from 40.3505 ng·hr/ml to 63.0476 ng·hr/ml, amean C_(max) for total estrone that is from 7.0858 ng/ml to 11.0715ng/ml, and a mean T_(max) for total estrone that is from 2 hr to 3.1 hr.

In some embodiments, method of treating a subject with a pharmaceuticalcomposition comprising estradiol and progesterone are provided. In someembodiments, the method comprises administering to the subject apharmaceutical composition comprising about 0.50 mg estradiol and about100 mg progesterone as described herein (e.g., a pharmaceuticalcomposition having the formulation of Formulation C in Table 3 above),wherein administration of the pharmaceutical composition produces, in aplasma sample from the subject, one or more parameters selected from: anAUC_((0-t)) for estradiol that is from 280.7467 pg·hr/ml to 438.6667pg·hr/ml; a C_(max) for estradiol that is from 12.9580 pg/ml to 20.2469pg/ml; a T_(max) for estradiol that is from 7.2 hr to 11.3 hr; anAUC_((0-t)) for progesterone that is from 48.0348 ng·hr/ml to 75.0543ng·hr/ml; a C_(max) for progesterone that is from 35.6889 ng/ml to55.7639 ng/ml; a T_(max) for progesterone that is from 2.4 hr to 3.8 hr;an AUC_((0-t)) for estrone that is from 1819.2181 pg·hr/ml to 2842.5283pg·hr/ml; a C_(max) for estrone that is from 85.3098 pg/ml to 133.2966pg/ml; a T_(max) for estrone that is from 4.4 hr to 6.9 hr; anAUC_((0-t)) for total estrone that is from 40.3505 ng·hr/ml to 63.0476ng·hr/ml; a C_(max) for total estrone that is from 7.0858 ng/ml to11.0715 ng/ml; and a T_(max) for total estrone that is from 2 hr to 3.1hr.

In some embodiments, the method further comprises obtaining a samplefrom the subject (e.g., a blood or plasma sample) followingadministration of a single dose of the pharmaceutical composition (e.g.,a pharmaceutical composition having the formulation of Formulation C inTable 3 above), and measuring one or more pharmacokinetic parametersselected from an AUC_((0-t)) for estradiol, a C_(max) for estradiol, anAUC_((0-t)) for progesterone, a C_(max) for progesterone, an AUC_((0-t))for estrone, a C_(max) for estrone, an AUC_((0-t)) for total estrone,and a C_(max) for total estrone; wherein the presence of one or more ofthe following values is indicative of a therapeutically effective dose:an AUC_((0-t)) for estradiol that is from 280.7467 pg·hr/ml to 438.6667pg·hr/ml; a C_(max) for estradiol that is from 12.9580 pg/ml to 20.2469pg/ml; an AUC_((0-t)) for progesterone that is from 48.0348 ng·hr/ml to75.0543 ng·hr/ml; a C_(max) for progesterone that is from 35.6889 ng/mlto 55.7639 ng/ml; an AUC_((0-t)) for estrone that is from 1819.2181pg·hr/ml to 2842.5283 pg·hr/ml; a C_(max) for estrone that is from85.3098 pg/ml to 133.2966 pg/ml; an AUC_((0-t)) for total estrone thatis from 40.3505 ng·hr/ml to 63.0476 ng·hr/ml; and a C_(max) for totalestrone that is from 7.0858 ng/ml to 11.0715 ng/ml. In some embodiments,the one or more pharmacokinetic parameters are measured about 18 hours,about 24 hours, about 18-36 hours, about 20-30 hours, about 22-26 hours,about 24-36 hours, about 36 hours, about 36-48 hours, about 40-48 hours,or about 48 hours after administration of the single dose.

AUC, C_(max), and T_(max) Parameters (D)

In some embodiments, a pharmaceutical composition of this disclosurecomprises estradiol at a dosage of about 1 mg and progesterone at adosage of about 100 mg. In some embodiments, the pharmaceuticalcomposition comprises the formulation of Formulation D in Table 4 above.

In some embodiments, administration of a composition comprising about 1mg estradiol and about 100 mg progesterone to a subject produces, in aplasma sample from the subject, one or both parameters selected from:

-   -   (i) an AUC_((0-t)) for estradiol that is from 561.4933 pg·hr/ml        to 877.3333 pg·hr/ml; or    -   (ii) a C_(max) for estradiol that is from 25.9161 pg/ml to        40.4939 pg/ml.

In some embodiments, administration of the composition to the subjectproduces both an AUC_((0-t)) for estradiol that is from 561.4933pg·hr/ml to 877.3333 pg·hr/ml, and a C_(max) for estradiol that is from25.9161 pg/ml to 40.4939 pg/ml.

In some embodiments, administration of the composition to the subjectfurther produces, in a plasma sample from the subject, one or bothparameters selected from:

-   -   (i) an AUC_((0-t)) for progesterone that is from 48.0348        ng·hr/ml to 75.0543 ng·hr/ml; or    -   (ii) a C_(max) for progesterone that is from 35.6889 ng/ml to        55.7639 ng/ml.

In some embodiments, administration of the composition to the subjectproduces both an AUC_((0-t)) for progesterone that is from 48.0348ng·hr/ml to 75.0543 ng·hr/ml, and a C_(max) for progesterone that isfrom 35.6889 ng/ml to 55.7639 ng/ml.

In some embodiments, administration of the composition to the subjectproduces, in a plasma sample from the subject,

-   -   (i) an AUC_((0-t)) for estradiol that is from 561.4933 pg·hr/ml        to 877.3333 pg·hr/ml;    -   (ii) a C_(max) for estradiol that is from 25.9161 pg/ml to        40.4939 pg/ml;    -   (iii) an AUC_((0-t)) for progesterone that is from 48.0348        ng·hr/ml to 75.0543 ng·hr/ml; or    -   (iv) a C_(max) for progesterone that is from 35.6889 ng/ml to        55.7639 ng/ml.

In some embodiments, administration of the composition to the subjectfurther produces, in a plasma sample from the subject, a T_(max) forestradiol that is from 7.2 hr to 11.3 hr. In some embodiments,administration of the composition to the subject further produces, in aplasma sample from the subject, a T_(max) for progesterone that is from2.4 hr to 3.8 hr.

In some embodiments, administration of the composition to the subjectproduces, in a plasma sample from the subject, one, two, three or moreparameters selected from:

-   -   (i) an AUC_((0-t)) for estradiol that is from 561.4933 pg·hr/ml        to 877.3333 pg·hr/ml;    -   (ii) a C_(max) for estradiol that is from 25.9161 pg/ml to        40.4939 pg/ml;    -   (iii) an AUC_((0-t)) for progesterone that is from 48.0348        ng·hr/ml to 75.0543 ng·hr/ml; or    -   (iv) a C_(max) for progesterone that is from 35.6889 ng/ml to        55.7639 ng/ml.

In some embodiments, administration of the pharmaceutical composition tothe subject produces both parameters (i) and (ii). In some embodiments,administration of the composition to the subject produces bothparameters (i) and (iii). In some embodiments, administration of thecomposition to the subject produces both parameters (i) and (iv). Insome embodiments, administration of the composition to the subjectproduces both parameters (ii) and (iii). In some embodiments,administration of the composition to the subject produces bothparameters (ii) and (iv). In some embodiments, administration of thecomposition to the subject produces both parameters (iii) and (iv). Insome embodiments, administration of the composition to the subjectproduces all of parameters (i), (ii), and (iii). In some embodiments,administration of the composition to the subject produces bothparameters (i), (iii), and (iv). In some embodiments, administration ofthe composition to the subject produces both parameters (ii), (iii), and(iv). In some embodiments, administration of the composition to thesubject produces all of parameters (i), (ii), (iii), and (iv).

In some embodiments, administration of the pharmaceutical composition tothe subject further produces, in a plasma sample from the subject, oneor more parameters selected from:

-   -   (i) an AUC_((0-t)) for estrone that is from 3638.4363 pg·hr/ml        to 5685.0567 pg·hr/ml;    -   (ii) a C_(max) for estrone that is from 170.6197 pg/ml to        266.5933 pg/ml; or    -   (iii) a T_(max) for estrone that is from 4.4 hr to 6.9 hr.

In some embodiments, administration of the pharmaceutical composition tothe subject further produces, in a plasma sample from the subject, oneor more parameters selected from:

-   -   (i) an AUC_((0-t)) for total estrone that is from 80.7010        ng·hr/ml to 126.0953 ng·hr/ml;    -   (ii) a C_(max) for total estrone that is from 14.1716 ng/ml to        22/1431 ng/ml; or    -   (iii) a T_(max) for total estrone that is from 2 hr to 3.1 hr.

In some embodiments, a pharmaceutical composition comprising about 1 mgestradiol and about 100 mg progesterone is administered to a populationof subjects in need thereof, and mean parameters are determined forsamples (e.g., blood or plasma samples) from the subjects administeredthe composition. Thus, in some embodiments, administration of thecomposition to a population of subject produces, in plasma samples fromthe subjects, one or more of a mean AUC_((0-t)) for estradiol that isfrom 561.4933 pg·hr/ml to 877.3333 pg·hr/ml, a mean C_(max) forestradiol that is from 25.9161 pg/ml to 40.4939 pg/ml, and a meanT_(max) for estradiol that is from 7.2 hr to 11.3 hr. In someembodiments, administration of the composition to a population ofsubject produces, in plasma samples from the subjects, one or more of amean AUC_((0-t)) for progesterone that is from 48.0348 ng·hr/ml to75.0543 ng·hr/ml, a mean C_(max) for progesterone that is from 35.6889ng/ml to 55.7639 ng/ml, and a mean T_(max) for progesterone that is from2.4 hr to 3.8 hr. In some embodiments, administration of the compositionto a population of subject produces, in plasma samples from thesubjects, one or more of a mean AUC_((0-t)) for estrone that is from3638.4363 pg·hr/ml to 5685.0567 pg·hr/ml, a mean C_(max) for estronethat is from 170.6197 pg/ml to 266.5933 pg/ml, and a mean T_(max) forestrone that is from 4.4 hr to 6.9 hr. In some embodiments,administration of the composition to a population of subject produces,in plasma samples from the subjects, one or more of a mean AUC_((0-t))for total estrone that is from 80.7010 ng·hr/ml to 126.0953 ng·hr/ml, amean C_(max) for total estrone that is from 14.1716 ng/ml to 22/1431ng/ml, and a mean T_(max) for total estrone that is from 2 hr to 3.1 hr.

In some embodiments, method of treating a subject with a pharmaceuticalcomposition comprising estradiol and progesterone are provided. In someembodiments, the method comprises administering to the subject apharmaceutical composition comprising about 1 mg estradiol and about 100mg progesterone as described herein (e.g., a pharmaceutical compositionhaving the formulation of Formulation D in Table 4 above), whereinadministration of the pharmaceutical composition produces, in a plasmasample from the subject, one or more parameters selected from: anAUC_((0-t)) for estradiol that is from 561.4933 pg·hr/ml to 877.3333pg·hr/m; a C_(max) for estradiol that is from 25.9161 pg/ml to 40.4939pg/ml; a T_(max) for estradiol that is from 7.2 hr to 11.3 hr; anAUC_((0-t)) for progesterone that is from 48.0348 ng·hr/ml to 75.0543ng·hr/ml; a C_(max) for progesterone that is from 35.6889 ng/ml to55.7639 ng/ml; a T_(max) for progesterone that is from 2.4 hr to 3.8 hr;an AUC_((0-t)) for estrone that is from 3638.4363 pg·hr/ml to 5685.0567pg·hr/ml; a C_(max) for estrone that is from 170.6197 pg/ml to 266.5933pg/ml; a T_(max) for estrone that is from 4.4 hr to 6.9 hr; anAUC_((0-t)) for total estrone that is from 80.7010 ng·hr/ml to 126.0953ng·hr/ml; a C_(max) for total estrone that is from 14.1716 ng/ml to22/1431 ng/ml; and a T_(max) for total estrone that is from 2 hr to 3.1hr.

In some embodiments, the method further comprises obtaining a samplefrom the subject (e.g., a blood or plasma sample) followingadministration of a single dose of the pharmaceutical composition (e.g.,a pharmaceutical composition having the formulation of Formulation D inTable 4 above), and measuring one or more pharmacokinetic parametersselected from an AUC_((0-t)) for estradiol, a C_(max) for estradiol, anAUC_((0-t)) for progesterone, a C_(max) for progesterone, an AUC_((0-t))for estrone, a C_(max) for estrone, an AUC_((0-t)) for total estrone,and a C_(max) for total estrone; wherein the presence of one or more ofthe following values is indicative of a therapeutically effective dose:an AUC_((0-t)) for estradiol that is from 561.4933 pg·hr/ml to 877.3333pg·hr/m; a C_(max) for estradiol that is from 25.9161 pg/ml to 40.4939pg/ml; an AUC_((0-t)) for progesterone that is from 48.0348 ng·hr/ml to75.0543 ng·hr/ml; a C_(max) for progesterone that is from 35.6889 ng/mlto 55.7639 ng/ml; an AUC_((0-t)) for estrone that is from 3638.4363pg·hr/ml to 5685.0567 pg·hr/ml; a C_(max) for estrone that is from170.6197 pg/ml to 266.5933 pg/ml; an AUC_((0-t)) for total estrone thatis from 80.7010 ng·hr/ml to 126.0953 ng·hr/ml; and a C_(max) for totalestrone that is from 14.1716 ng/ml to 22/1431 ng/ml. In someembodiments, the one or more pharmacokinetic parameters are measuredabout 18 hours, about 24 hours, about 18-36 hours, about 20-30 hours,about 22-26 hours, about 24-36 hours, about 36 hours, about 36-48 hours,about 40-48 hours, or about 48 hours after administration of the singledose.

AUC, C_(max), and T_(max) Parameters (E)

In some embodiments, a pharmaceutical composition of this disclosurecomprises estradiol at a dosage of about 2 mg and progesterone at adosage of about 200 mg. In some embodiments, the pharmaceuticalcomposition comprises the formulation of Formulation E in Table 5 above.

In some embodiments, administration of a pharmaceutical compositioncomprising about 2 mg estradiol and about 200 mg progesterone to asubject produces, in a plasma sample from the subject, one or bothparameters selected from:

-   -   (i) an AUC_((0-t)) for estradiol that is from 1123 pg·h/ml to        1755 pg·h/ml; or    -   (ii) a C_(max) for estradiol that is from 52 pg/ml to 81 pg/ml.

In some embodiments, administration of the composition to the subjectproduces both an AUC_((0-t)) for estradiol that is from 1123 pg·h/ml to1755 pg·h/ml, and a C_(max) for estradiol that is from 52 pg/ml to 81pg/ml.

In some embodiments, administration of the composition to the subjectfurther produces, in a plasma sample from the subject, one or bothparameters selected from:

-   -   (i) an AUC_((0-t)) for progesterone that is from 96 ng·hr/ml to        150 ng·hr/ml; or    -   (ii) a C_(max) for progesterone that is from 71 ng/ml to 112        ng/ml.

In some embodiments, administration of the composition to the subjectproduces both an AUC_((0-t)) for progesterone that is from 96 ng·hr/mlto 150 ng·hr/ml, and a C_(max) for progesterone that is from 71 ng/ml to112 ng/ml.

In some embodiments, administration of the composition to the subjectproduces, in a plasma sample from the subject,

-   -   (i) an AUC_((0-t)) for estradiol that is from 1123 pg·h/ml to        1755 pg·h/ml;    -   (ii) a C_(max) for estradiol that is from 52 pg/ml to 81 pg/ml;    -   (iii) an AUC_((0-t)) for progesterone that is from 96 ng·hr/ml        to 150 ng·hr/ml; or    -   (iv) a C_(max) for progesterone that is from 71 ng/ml to 112        ng/ml.

In some embodiments, administration of the composition to the subjectfurther produces, in a plasma sample from the subject, a T_(max) forestradiol that is from 7.2 hr to 11.3 hr. In some embodiments,administration of the composition to the subject further produces, in aplasma sample from the subject, a T_(max) for progesterone that is from2.4 hr to 3.8 hr.

In some embodiments, administration of the pharmaceutical composition tothe subject produces, in a plasma sample from the subject, one, two,three or more parameters selected from:

-   -   (i) an AUC_((0-t)) for estradiol that is from 1123 pg·h/ml to        1755 pg·h/ml;    -   (ii) a C_(max) for estradiol that is from 52 pg/ml to 81 pg/ml;    -   (iii) an AUC_((0-t)) for progesterone that is from 96 ng·hr/ml        to 150 ng·hr/ml; or    -   (iv) a C_(max) for progesterone that is from 71 ng/ml to 112        ng/ml.

In some embodiments, administration of the pharmaceutical composition tothe subject produces both parameters (i) and (ii). In some embodiments,administration of the composition to the subject produces bothparameters (i) and (iii). In some embodiments, administration of thecomposition to the subject produces both parameters (i) and (iv). Insome embodiments, administration of the composition to the subjectproduces both parameters (ii) and (iii). In some embodiments,administration of the composition to the subject produces bothparameters (ii) and (iv). In some embodiments, administration of thecomposition to the subject produces both parameters (iii) and (iv). Insome embodiments, administration of the composition to the subjectproduces all of parameters (i), (ii), and (iii). In some embodiments,administration of the composition to the subject produces bothparameters (i), (iii), and (iv). In some embodiments, administration ofthe composition to the subject produces both parameters (ii), (iii), and(iv). In some embodiments, administration of the composition to thesubject produces all of parameters (i), (ii), (iii), and (iv).

In some embodiments, administration of the pharmaceutical composition tothe subject further produces, in a plasma sample from the subject, oneor more parameters selected from:

-   -   (i) an AUC_((0-t)) for estrone that is from 7277 pg·hr/ml to        11370 pg·hr/ml;    -   (ii) a C_(max) for estrone that is from 341 pg/ml to 533 pg/ml;        or    -   (iii) a T_(max) for estrone that is from 4.4 hr to 6.9 hr.

In some embodiments, administration of the pharmaceutical composition tothe subject further produces, in a plasma sample from the subject, oneor more parameters selected from:

-   -   (i) an AUC_((0-t)) for total estrone that is from 161 ng·h/ml to        252 ng·h/ml    -   (ii) a C_(max) for total estrone that is from 28 ng/ml to 44        ng/ml; or    -   (iii) a T_(max) for total estrone that is from 2 hr to 3.1 hr.

In some embodiments, a pharmaceutical composition comprising about 2 mgestradiol and about 200 mg progesterone is administered to a populationof subjects in need thereof, and mean parameters are determined forsamples (e.g., blood or plasma samples) from the subjects administeredthe composition. Thus, in some embodiments, administration of thecomposition to a population of subject produces, in plasma samples fromthe subjects, one or more of a mean AUC_((0-t)) for estradiol that isfrom 1123 pg·h/ml to 1755 pg·h/ml, a mean C_(max) for estradiol that isfrom 52 pg/ml to 81 pg/ml, and a mean T_(max) for estradiol that is from7.2 hr to 11.3 hr. In some embodiments, administration of thecomposition to a population of subject produces, in plasma samples fromthe subjects, one or more of a mean AUC_((0-t)) for progesterone that isfrom 96 ng·hr/ml to 150 ng·hr/ml, a mean C_(max) for progesterone thatis from 71 ng/ml to 112 ng/ml, and a mean T_(max) for progesterone thatis from 2.4 hr to 3.8 hr. In some embodiments, administration of thecomposition to a population of subject produces, in plasma samples fromthe subjects, one or more of a mean AUC_((0-t)) for estrone that is from7277 pg·hr/ml to 11370 pg·hr/ml, a mean C_(max) for estrone that is from341 pg/ml to 533 pg/ml, and a mean T_(max) for estrone that is from 4.4hr to 6.9 hr. In some embodiments, administration of the composition toa population of subject produces, in plasma samples from the subjects,one or more of a mean AUC_((0-t)) for total estrone that is from 161ng·h/ml to 252 ng·h/ml, a mean C_(max) for total estrone that is from 28ng/ml to 44 ng/ml, and a mean T_(max) for total estrone that is from 2hr to 3.1 hr.

In some embodiments, method of treating a subject with a pharmaceuticalcomposition comprising estradiol and progesterone are provided. In someembodiments, the method comprises administering to the subject apharmaceutical composition comprising about 2 mg estradiol and about 200mg progesterone as described herein (e.g., a pharmaceutical compositionhaving the formulation of Formulation E in Table 5 above), whereinadministration of the pharmaceutical composition produces, in a plasmasample from the subject, one or more parameters selected from: anAUC_((0-t)) for estradiol that is from 1123 pg·h/ml to 1755 pg·h/ml; aC_(max) for estradiol that is from 52 pg/ml to 81 pg/ml; a T_(max) forestradiol that is from 7.2 hr to 11.3 hr; an AUC_((0-t)) forprogesterone that is from 96 ng·hr/ml to 150 ng·hr/ml; a C_(max) forprogesterone that is from 71 ng/ml to 112 ng/ml; a T_(max) forprogesterone that is from 2.4 hr to 3.8 hr; an AUC_((0-t)) for estronethat is from 7277 pg·hr/ml to 11370 pg·hr/ml; a C_(max) for estrone thatis from 341 pg/ml to 533 pg/ml; a T_(max) for estrone that is from 4.4hr to 6.9 hr; an AUC_((0-t)) for total estrone that is from 161 ng·h/mlto 252 ng·h/ml; a C_(max) for total estrone that is from 28 ng/ml to 44ng/ml; and a T_(max) for total estrone that is from 2 hr to 3.1 hr.

In some embodiments, the method further comprises obtaining a samplefrom the subject (e.g., a blood or plasma sample) followingadministration of a single dose of the pharmaceutical composition (e.g.,a pharmaceutical composition having the formulation of Formulation E inTable 5 above), and measuring one or more pharmacokinetic parametersselected from an AUC_((0-t)) for estradiol, a C_(max) for estradiol, anAUC_((0-t)) for progesterone, a C_(max) for progesterone, an AUC_((0-t))for estrone, a C_(max) for estrone, an AUC_((0-t)) for total estrone,and a C_(max) for total estrone; wherein the presence of one or more ofthe following values is indicative of a therapeutically effective dose:an AUC_((0-t)) for estradiol that is from 1123 pg·h/ml to 1755 pg·h/ml;a C_(max) for estradiol that is from 52 pg/ml to 81 pg/ml; anAUC_((0-t)) for progesterone that is from 96 ng·hr/ml to 150 ng·hr/ml; aC_(max) for progesterone that is from 71 ng/ml to 112 ng/ml; anAUC_((0-t)) for estrone that is from 7277 pg·hr/ml to 11370 pg·hr/ml; aC_(max) for estrone that is from 341 pg/ml to 533 pg/ml; an AUC_((0-t))for total estrone that is from 161 ng·h/ml to 252 ng·h/ml; and a C_(max)for total estrone that is from 28 ng/ml to 44 ng/ml. In someembodiments, the one or more pharmacokinetic parameters are measuredabout 18 hours, about 24 hours, about 18-36 hours, about 20-30 hours,about 22-26 hours, about 24-36 hours, about 36 hours, about 36-48 hours,about 40-48 hours, or about 48 hours after administration of the singledose.

In some embodiments, administration of the pharmaceutical composition asdescribed herein results in the blood plasma estradiol concentrationprofile of FIG. 1. In some embodiments, administration of thepharmaceutical composition results in the blood plasma progesteroneconcentration profile of FIG. 2. In some embodiments, administration ofthe pharmaceutical composition results in the blood plasma estroneconcentration profile of FIG. 3. In some embodiments, administration ofthe pharmaceutical composition results in the blood plasma total estroneconcentration profile of FIG. 4.

Administration and Treatment

Pharmaceutical compositions comprising estradiol and progesterone asdescribed herein (e.g., compositions comprising solubilized estradiol,suspended progesterone, and a medium chain solubilizing agent) can beprepared and administered in a wide variety of oral, parenteral andtopical dosage forms. Oral preparations include tablets, pills, powder,dragees, capsules, liquids, lozenges, cachets, gels, syrups, slurries,suspensions, etc., suitable for ingestion by the patient. Pharmaceuticalcompositions can be formulated for any appropriate manner ofadministration, including, for example, topical, oral, nasal,intrathecal, rectal, vaginal, sublingual or parenteral administration,including subcutaneous, intravenous, intramuscular, intrasternal,intracavernous, intrameatal, or intraurethral injection or infusion. Insome embodiments, administration is by injection, that is,intravenously, intramuscularly, intracutaneously, subcutaneously,intraduodenally, or intraperitoneally.

For preparing pharmaceutical compositions from the compounds of thisdisclosure, the pharmaceutically acceptable compositions can be eithersolid or liquid. Solid form preparations include powders, tablets,pills, capsules, cachets, suppositories, and dispersible granules. Asolid preparation can comprise one or more substances, which may alsoact as diluents, flavoring agents, binders, preservatives, tabletdisintegrating agents, or an encapsulating material. Details ontechniques for formulation and administration are well described in thescientific and patent literature, see, e.g., the latest edition ofRemington's Pharmaceutical Sciences, Mack Publishing Co, Easton Pa.(“Remington's”).

In general, the type of composition is selected based on the mode ofadministration. A pharmaceutical composition (e.g., for oraladministration or delivery by injection) can be in the form of a liquid(e.g., an elixir, syrup, solution, emulsion or suspension).Alternatively, a pharmaceutical composition as described herein can takethe form of a pill, tablet, or capsule containing the liquid oil, andthus, the composition can contain any of the following: a diluent suchas lactose, sucrose, dicalcium phosphate, and the like; a disintegrantsuch as starch or derivatives thereof; a lubricant such as magnesiumstearate and the like; and a binder such a starch, gum acacia,polyvinylpyrrolidone, gelatin, cellulose and derivatives thereof. Thecomposition can also be formulated into a suppository disposed, forexample, in a polyethylene glycol (PEG) solubilizing agent.

Administration of the compositions of this disclosure can be carried outvia any of the accepted modes of administration. Thus, administrationcan be, for example, intravenous, topical, subcutaneous, transcutaneous,transdermal, intramuscular, oral, intra-joint, parenteral,intra-arteriole, intradermal, intraventricular, intracranial,intraperitoneal, intralesional, intranasal, rectal, vaginal, or byinhalation. In some embodiments, a composition as described herein isadministered orally. For example, a pharmaceutical composition asdescribed herein can be administered via capsules such as soft capsules.

In some embodiments, a pharmaceutical composition as described herein isadministered once daily for a period of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 60, 70,80, 90, 100 days or more. In some embodiments, a pharmaceuticalcomposition as described herein is administered daily for at least oneweek, at least two weeks, at least three weeks, at least four weeks, atleast one month, at least two months, at least three months, at leastfour months, at least five months, at least six months, at least sevenmonths, at least eight months, at least nine months, at least tenmonths, at least eleven months, at least twelve months, or more. In someembodiments, a pharmaceutical composition as described herein isadministered as a continuous-combined therapy regimen.

In some embodiments, a 28-day or monthly regimen of daily doses ispackaged in a single kit (e.g., a blister pack) having administrationdays identified to improve compliance and reduce associated symptoms,among others. In some embodiments, each daily dose contains bothestradiol and progesterone. In some embodiments, one or more of thedaily doses contains no estradiol or no progesterone. Daily doses thatcomprise no estradiol or progesterone API may be referred to asplacebos. A blister pack can have a plurality of scores or perforationsseparating the blister pack into 28 days. Each day may further comprisea single blister or a plurality of blisters. In various embodiments,each unit dose may contain micronized or partially solubilized, or fullysolubilized progesterone or solubilized estradiol in amounts as setforth herein, although other dose ranges may be contemplated. Inaddition, kits having other configurations are also contemplated herein.For example, without limitation, kits having such blister packs maycontain any number of daily doses.

In some embodiments, the pharmaceutical compositions disclosed hereinare useful in treating conditions in subjects caused, at least in part,by estrogen deficiency, particularly for women with a uterus. Forexample, in embodiments, the pharmaceutical compositions disclosedherein are useful for the treatment of one or more of the followingconditions: endometrial hyperplasia; secondary amenorrhea; prevention ofpreterm birth, when the subject has a shortened cervix;menopause-related symptoms including, for example, vasomotor symptoms;in relation to treatment of hypoestrogenism related symptoms including,for example and without limitation, hot flashes and night sweats(vasomotor symptoms), sleep disturbances, mood changes and vulvo-vaginalatrophy; and osteoporosis and other non-menopausal disease states orconditions treated with supplemental progesterone or estrogen. In someembodiments, the pharmaceutical compositions disclosed herein are usefulin treating vasomotor symptoms, including but not limited to, hotflashes and night sweats. In some embodiments, the pharmaceuticalcompositions disclosed herein are useful in treating hot flashes andnight sweats.

In some embodiments, the pharmaceutical compositions disclosed hereinare useful in treating hot flashes. Thus, in some embodiments, thisdisclosure provides methods of treating such a condition byadministering to the subject a composition comprising estradiol andprogesterone as described herein.

III. EXAMPLES

The following examples are offered to illustrate, but not to limit, theclaimed subject matter.

Example 1

In an exemplary embodiment, a soft gelatin capsule contains apharmaceutical composition comprising suspended progesterone andsolubilized estradiol:

TABLE 6 Qty/Capsule Ingredient Mass (mg) % w/w (mg) Progesterone, USP,micronized 50.00 7.14 50.00 Estradiol Hemihydrate, USP 2.03 0.29 2.03CAPMUL ® MCM, NF 82.57 577.97 GELUCIRE ® 44/14, NF 10.0 70.00 TOTAL100.00 700.00

The encapsulated pharmaceutical composition of Table 6 may bemanufactured in any suitable manner. For the purposes of this Example,mixing may be facilitated by an impellor, agitator, or other suitablemeans. Also for the purposes of this Example, heating or mixing may beperformed under an inert or relatively inert gas atmosphere, such asnitrogen gas (N₂). Mixing or heating for the purposes of this Examplemay be performed in any suitable vessel, such as a stainless steelvessel.

For example, CAPMUL® MCM may be heated to between 30° C. to 50° C., morepreferably from 35° C. to 45° C., and more preferably to 40° C.±2° C.GELUCIRE® 44/14 may be added to the CAPMUL® MCM and mixed untildissolved (to increase the solubility of progesterone in the finalsolution, GELUCIRE® 44/14 was added at about 10% w/w). The addition mayoccur all at once or may occur gradually over a period of time. Heat maycontinue to be applied during the mixing of the GELUCIRE® 44/14 and theCAPMUL® MCM.

Heat may be removed from the GELUCIRE® 44/14 and CAPMUL® MCM mixture.Estradiol Hemihydrate may be added to the mixture. The addition mayoccur all at once or may occur gradually over a period of time.Micronized progesterone may then be added to the GELUCIRE® 44/14,CAPMUL® MCM and Estradiol Hemihydrate mixture until dissolved.

The addition may occur all at once or may occur gradually over a periodof time.

Example 2

An example of the final scale-up formulation is provided in Table 7. Tomanufacture, CAPMUL® MCM is heated to 40° C. GELUCIRE® 44/14 is heatedto 65° C. and added and mixed until dissolved. Heat is removed.Estradiol is added and mixed until dissolved. Micronized progesterone isthen added and mixed until fully suspended.

TABLE 7 Quantitative Formula: Batch Size 10,000 capsules Label Qty/Amount/ Item Claim Capsule Batch No. Ingredient (mg) % w/w (mg) (kg) 1.Progesterone, USP, 50.00 7.14 50.00 0.50 micronized 2. EstradiolHemihydrate, 2.03 0.29 2.03 0.02 USP 3. CAPMUL ® MCM, NF 82.57 577.975.78 4. GELUCIRE ® 44/14, 10.0 70.00 0.70 NF Total: 100.00 700.00 7.00

Example 3

In an exemplary embodiment, a soft gelatin capsule contains apharmaceutical composition having fully solubilized estradiol andpartially solubilized progesterone comprising:

TABLE 8 Label Qty/ Amount/ Item Claim Capsule Batch No. Ingredient (mg)% w/w (mg) (g) 1. Progesterone, USP, 50.00 25.000 50.00 500.00micronized 2. Estradiol Hemihydrate 0.25 0.129 0.26 2.58 3. CAPMUL ®MCM, NF 73.371 146.74 1467.42 4. GELUCIRE ® 44/14, 1.500 3.00 30.00 NFTotal: 100.000 200.00 mg 2000.00

To manufacture, CAPMUL® MCM is heated to 65° C. GELUCIRE® 44/14 is addedand mixed until dissolved. Heat is removed. Estradiol is added and mixeduntil dissolved. Micronized progesterone is then added and dispersed.The mixture is then passed through a colloid mill. The resultant fillmass can be used for encapsulation.

Example 4

In an exemplary embodiment, a soft gelatin capsule contains apharmaceutical composition having fully solubilized estradiol andpartially solubilized progesterone comprising:

TABLE 9 Label Qty/ Amount/ Item Claim Capsule Batch No. Ingredient (mg)% w/w (mg) (g) 1. Progesterone, USP, 200.00 33.33 200.0 2000.0micronized 2. Estradiol Hemihydrate 2.00 0.35 2.07 20.7 3. CAPMUL ® MCM,NF 65.32 391.93 3919.3 4. GELUCIRE ® 44/14, 1.00 6.0 60.0 NF Total:100.00 600.0 mg 6000.0

To manufacture, CAPMUL® MCM is heated to 65° C. GELUCIRE® 44/14 is addedand mixed until dissolved. Heat is removed. Estradiol is added and mixeduntil dissolved. Micronized progesterone is then added and dispersed.The mixture is then passed through a colloid mill. The resultingpharmaceutical composition is encapsulated in soft gelatin capsules.Alternatively, GELUCIRE® 44/14 is heated to 65° C. and CAPMUL® MCM isheated to 40° C.±5° C. to achieve mixing of the oil and the surfactantbefore heat is removed; estradiol is added while the mixture is cooling;progesterone is added when the mixture has dropped below about 40° C.;the mixture is then passed through a colloid mill one or more times,e.g., three times.

Example 5

Pharmacokinetics of the First Combination 17β-Estradiol/ProgesteroneCapsule in Clinical Development for Hormone Therapy

The objective of this study was to evaluate the pharmacokinetic and oralbioavailability of a combination capsule of 17β-estradiol/progesteronein comparison to co-administration of the individual products ESTRACE®and PROMETRIUM®.

Subjects and Study Design: An open label, balanced, randomized,single-dose, 2-treatment, 3-period, 3-sequence, crossover,partial-replicate, reference-scaled, oral, relative bioavailabilitystudy compared the bioavailability of an investigational 2-mg17β-estradiol/200-mg progesterone combination capsule, without peanutoil (formulated in a manner similar to that set forth in Table 9), withthat of co-administered 200-mg PROMETRIUM® (progesterone) and 2-mgESTRACE® (17β-estradiol) tablets in healthy postmenopausal women aged40-65 years (N=66). Key inclusion criteria for subjects included a BMI18.50 to 29.99 kg/m² who were nonsmokers or ex-smokers (no smoking inthe last 3 months). Key exclusion criteria for subjects includedconsuming grapefruit juice or poppy-containing foods within 48 hoursbefore and throughout the study, use of any hormonal agent within 14days before the study, and use of menopausal hormone therapy within 6months before dosing.

Patients were randomly assigned sequentially to 1 of 3 dosing sequencesof the same dose of the combination capsule (Test, T) and referenceproducts (Reference, R): TRR, RTR, or RRT. 66 subjects were randomizedand 62 (94.0%) completed the study. Subjects had a mean age of 49.5±5.6years (range 40 to 64) and a mean BMI of 24.8±3.1 kg/m² (range18.7-29.9).

After consuming a high-fat, high-calorie breakfast, each woman receiveda single dose of the combination (Test) capsule in 1 period of the studyand single doses of the co-administered products (Reference) in each ofthe 2 remaining periods. Blood samples were collected within 75 minutesbefore dosing and post-dose at 0.25, 0.5, 0.67, 0.83, 1, 1.33, 1.67, 2,2.5, 3, 4, 5, 6, 7, 8, 10, 12, 18, 24, 36, and 48 hours after dosing todetermine progesterone, free (unconjugated) estradiol, and free andtotal (conjugated+free, including estrone sulfates) estroneconcentrations. After collection of blood samples at each time point,the blood samples were centrifuged at 4000 RPM for 10 minutes at 4° C.to separate the plasma. The plasma from samples was separated into twoaliquots. 1.5 mL from the plasma sample was transferred into aliquot I,and the remaining plasma sample was transferred into aliquot II. Thesealiquots were stored at −30° C. for interim storage, then at −70° C.until completion of the analysis.

Progesterone, estradiol, estrone, and total estrone in human plasma wasdetermined using the LC-MS/MS method. The primary (C_(max), AUC_(0-t),and AUC_(0-∞)) and secondary (T_(max), t_(1/2), and K_(e)) PK parametersfor each analyte were determined for each subject during each period bynon-compartment analyses using baseline-adjusted concentrations.Statistical analyses were conducted using the SAS® statistical software.

Results: The mean, standard deviation (SD), geometric mean, coefficientof variation (CV %), minimum, median, and maximum were calculated forC_(max), AUC_(0-t), AUC_(0-∞), T_(max), t_(1/2), K_(el), K_(el_lower),K_(el_Upper), and AUC_(% Extrap_obs) for progesterone, estradiol,estrone, and total estrone. The results are presented in Tables 10, 11,12, and 13 below. For each of Tables 10-13, “Test Product (T)” refers tothe progesterone+estradiol pharmaceutical composition, while “Referenceproduct (R1)” and “Reference product (R2)” refers to co-administeredPROMETRIUM® (progesterone) and ESTRACE® (estradiol). Blood plasmaconcentrations of progesterone, estradiol, estrone, and total estroneover time are also shown in FIGS. 1-4.

TABLE 10 Summary of Pharmacokinetic Parameters of Test Product (T)versus Reference Product (R₁, R₂) for Progesterone Untransformed Data(Mean ± SD) PK Parameter N Test Product (T) N Reference product (R1) NReference product (R2) C_(max) (ng/mL) 62  89.2222 ± 149.7309 62 72.7228 ± 101.8885 62 69.7590 ± 87.0777 AUC_(0-t) (ng · hr/mL) 62120.0869 ± 164.1385 62 125.9406 ± 152.3483 62 111.5867 ± 113.3200AUC_(0-∞) (ng · hr/mL) 57 131.3817 ± 172.4806 57 142.1332 ± 160.4853 56126.6006 ± 117.2665 T_(max) (hr) 62 3.00(0.83-10.00) 62 3.00(1.00-12.00)62 4.00(0.67-18.00) K_(el) (hr⁻¹) 57 0.3064 ± 0.2427 57 0.2684 ± 0.191256 0.2795 ± 0.2475 t_(1/2) (hr) 57 4.6445 ± 4.5366 57 5.1555 ± 4.9794 565.0389 ± 4.5887 K_(el)_ Lower (hr⁻¹) 57 7.6667 ± 4.6047 57 7.4123 ±4.2164 56 7.9018 ± 3.9120 K_(el)_Upper (hr⁻¹) 57 16.2218 ± 11.0051 5719.1728 ± 12.3801 56 18.1975 ± 10.0858 AUC_Extra (%) 57 4.3374 ± 2.552857 4.8416 ± 3.7526 56 5.1868 ± 4.1434 *Expressed in terms of median(range)

TABLE 11 Summary of Pharmacokinetic Parameters of Test Product (T)versus Reference Product (R₁, R₂) for Estradiol Untransformed Data (Mean± SD) PK Parameter Test Product (T) Reference product (R1) Referenceproduct (R2) Cmax (pg/mL) 64.7902 ± 50.9833 69.1286 ± 33.0484 73.4236 ±43.4077 AUC_(0-t) (pg · hr/mL) 1403.7333 ± 763.8136  1508.2206 ±876.7390  1658.2502 ± 976.5556  AUC_(0-∞) (pg · hr/mL) 2459.4394 ±4498.2737 2842.8805 ± 4582.6502 2110.9591 ± 1175.3995 T_(max) (hr)9.00(0.50-36.00) 10.0(0.50-35.12) 10.00(0.25-36.60) K_(el) (hr⁻¹)0.04380.0197 0.0457 ± 0.0358 0.0464 ± 0.0338 t_(1/2) (hr) 31.9104 ±95.9769 25.0908 ± 28.8346 20.8774 ± 12.0825 K_(el)_Lower (hr⁻¹) 14.9472± 7.2715  14.9667 ± 7.0150  14.7953 ± 5.8774  K_(el)_Upper (hr⁻¹)45.3602 ± 6.3668  44.3277 ± 7.4003  43.8330 ± 7.6449  AUC_Extra (%)22.8106 ± 16.6498 25.4773 ± 20.2911 24.9566 ± 16.4713 *Expressed interms of median (range)

TABLE 12 Summary of Pharmacokinetic Parameters of Test Product (T)versus Reference Product (R₁, R₂) for Free Estrone Untransformed Data(Mean ± SD) PK Parameter Test Product (T) Reference product (R1)Reference product (R2) Cmax (pg/mL)  426.5492 ± 179.3303 455.5107 ±189.448  467.2302 ± 207.4373 AUC_(0-t) (pg · hr/mL)  9096.0907 ±4377.2730 10156.0282 ± 5140.5831 10507.3557 ± 5183.1289 AUC_(0-∞) (pg ·hr/mL) 11994.9695 ± 6678.5468 13445.9048 ± 8699.4068 14066.2362 ±7563.2370 T_(max) (hr) 5.50(0.83-36.00) 8.00(1.67-18.00)10.00(1.67-18.00) K_(el) (hr⁻¹)  0.0399 ± 0.0146  0.0424 ± 0.0172 0.0406 ± 0.0209 t_(1/2) (hr) 20.3172 ± 9.4052 19.4595 ± 9.8711 20.7515± 9.3985 K_(el)_Lower (hr⁻¹) 13.8443 ± 7.0649 14.8871 ± 6.6459 14.9194 ±6.4485 K_(el)_Upper (hr⁻¹) 46.0238 ± 5.5080 46.2547 ± 5.3060 46.2244 ±5.3126 AUC_Extra (%)  21.2980 ± 11.2283  20.3648 ± 11.1060  21.8900 ±11.8537 *Expressed in terms of median (range)

TABLE 13 Summary of Pharmacokinetic Parameters of Test Product (T)versus Reference Product (R₁, R₂) for Total Estrone Untransformed Data(Mean ± SD) PK Parameter N Test Product (T) N Reference product (R1) NReference product (R2) C_(max) (ng/mL) 61 35.4289 ± 17.0856 61 19.8716 ±7.4485  61 19.9048 ± 8.0288  AUC_(0-t) (ng · hr/mL) 61 201.7524 ±94.2081  61 182.7729 ± 88.8386  61 199.8295 ± 94.9392  AUC_(0-∞) (ng ·hr/mL) 61 213.2402 ± 104.6011 60 193.6387 ± 100.5831 56 203.0289 ±81.4884  T_(max) (hr) 61 2.50(0.67-7.00) 61 4.00(1.33-18.00) 614.00(1.33-10.00) K_(el) (hr⁻¹) 61 0.0799 ± 0.0398 60 0.0803 ± 0.0399 560.0718 ± 0.0243 t_(1/2) (hr) 61 10.3619 ± 4.0023  60 9.8448 ± 3.0702 5610.7830 ± 3.6624  K_(el)_ Lower (hr⁻¹) 61 13.0492 ± 6.8585  60 13.5945 ±8.0129  56 11.8870 ± 6.8696  K_(el)_Upper (hr⁻¹) 61 45.3979 ± 6.6589  6046.3775 ± 5.2525  56 46.7054 ± 4.3888  AUC_Extra (%) 61 4.5030 ± 3.736660 4.5913 ± 3.4953 56 5.3450 ± 3.9831 *Expressed in terms of median(range)

Example 6

Pharmacokinetic data (C_(max), AUC_((0-t)), AUC_((0-∞)), and T_(max))for progesterone, estradiol, free estrone, and total estrone ispresented in Tables 14-17. Pharmaceutical compositions A-E are disclosedin Tables 1-5. The pK values for pharmaceutical composition E werecalculated as disclosed in Example 5. For pharmaceutical compositionsA-D, expected pharmacokinetic data is calculated from the data disclosedfor pharmaceutical composition E.

TABLE 14 Summary of Pharmacokinetic Parameters of the PharmaceuticalCompositions of Tables 1-5 for Progesterone Pharmaceutical ProgesteroneEstradiol C_(max) AUC_((0-t)) AUC_((0-∞)) T_(max) Composition ContentContent (ng/mL) (ng · hr/mL) (ng · hr/mL) (hr) A  50 mg 0.25 mg 22.3055530.0217 32.8454 3.00 B  50 mg 0.50 mg 22.3055 30.0217 32.8454 3.00 C 100mg 0.50 mg 44.6111 60.0435 65.6909 3.00 D 100 mg   1 mg 44.6111 60.043565.6909 3.00 E 200 mg   2 mg 89.2222 120.0869 131.3817 3.00

TABLE 15 Summary of Pharmacokinetic Parameters of the PharmaceuticalCompositions of Tables 1-5 for Estradiol Pharmaceutical ProgesteroneEstradiol C_(max) AUC_((0-t)) AUC_((0-∞)) T_(max) Composition ContentContent (pg/mL) (pg · hr/mL) (pg · hr/mL) (hr) A  50 mg 0.25 mg 8.0988175.4667 307.4299 9.00 B  50 mg 0.50 mg 16.1976 350.9333 614.8599 9.00 C100 mg 0.50 mg 16.1976 350.9333 614.8599 9.00 D 100 mg   1 mg 32.3951701.8667 1229.7197 9.00 E 200 mg   2 mg 64.7902 1403.7333 2459.4394 9.00

TABLE 16 Summary of Pharmacokinetic Parameters of the PharmaceuticalCompositions of Tables 1-5 for Free Estrone Pharmaceutical ProgesteroneEstradiol C_(max) AUC_((0-t)) AUC_((0-∞)) T_(max) Composition ContentContent (pg/mL) (pg · hr/mL) (pg · hr/mL) (hr) A  50 mg 0.25 mg 53.31871137.0113 1499.3712 5.50 B  50 mg 0.50 mg 106.6373 2274.0227 2998.74245.50 C 100 mg 0.50 mg 106.6373 2274.0227 2998.7424 5.50 D 100 mg   1 mg213.2746 4548.0454 5997.4848 5.50 E 200 mg   2 mg 426.5492 9096.090711994.9695 5.50

TABLE 17 Summary of Pharmacokinetic Parameters of the PharmaceuticalCompositions of Tables 1-5 for Total Estrone Pharmaceutical ProgesteroneEstradiol C_(max) AUC_((0-t)) AUC_((0-∞)) T_(max) Composition ContentContent (ng/mL) (ng · hr/mL) (ng · hr/mL) (hr) A  50 mg 0.25 mg 4.428625.2191 26.6550 2.50 B  50 mg 0.50 mg 8.8572 50.4381 53.3101 2.50 C 100mg 0.50 mg 8.8572 50.4381 53.3101 2.50 D 100 mg   1 mg 17.7145 100.8762106.6201 2.50 E 200 mg   2 mg 35.4289 201.7524 213.2402 2.50

The ranges of expected pK values for each of the pharmaceuticalcompositions of Tables 1-4 are disclosed in Tables 18-21, respectively.

TABLE 18 pK Ranges for the Pharmaceutical Composition of Table 1(Pharmaceutical Composition A) C_(max) AUC_((0-t)) AUC_((0-∞))Progesterone 17.8444 ng/mL to 24.0174 ng · hr/mL to 26.2763 ng · hr/mLto 27.8819 ng/mL 37.5272 ng · hr/mL 41.0568 ng · hr/mL Estradiol 6.4790pg/mL to 140.3733 pg · hr/mL to 245.9439 pg · hr/mL to 10.1235 pg/mL219.3333 pg · hr/mL 384.2874 pg · hr/mL Free estrone 42.6549 pg/mL to909.6091 pg · hr/mL to 1199.4970 pg · hr/mL to 66.6483 pg/mL 1421.2642pg · hr/mL 1874.2140 pg · hr/mL Total estrone 3.5429 ng/mL to 20.1752 ng· hr/mL to 21.3240 ng · hr/mL to 5.5358 ng/mL 31.5238 ng · hr/mL 33.3188ng · hr/mL

TABLE 19 pK Ranges for the Pharmaceutical Composition of Table 2(Pharmaceutical Composition B) C_(max) AUC_((0-t)) AUC_((0-∞))Progesterone 17.8444 ng/mL to 24.0174 ng · hr/mL to 26.2763 ng · hr/mLto 27.8819 ng/mL 37.5272 ng · hr/mL 41.0568 ng · hr/mL Estradiol 12.9580pg/mL to 280.7467 pg · hr/mL to 491.8879 pg · hr/mL to 20.2469 pg/mL438.6667 pg · hr/mL 768.5748 pg · hr/mL Free estrone 85.3098 pg/mL to1819.2181 pg · hr/mL to 2398.9939 pg · hr/mL to 133.2966 pg/mL 2842.5283pg · hr/mL 3748.4280 pg · hr/mL Total estrone 7.0858 ng/mL to 40.3505 ng· hr/mL to 42.6480 ng · hr/mL to 11.0715 ng/mL 63.0476 ng · hr/mL66.6376 ng · hr/mL

TABLE 20 pK Ranges for the Pharmaceutical Composition of Table 3(Pharmaceutical Composition C) C_(max) AUC_((0-t)) AUC_((0-∞))Progesterone 35.6889 ng/mL to 48.0348 ng · hr/mL to 52.5527 ng · hr/mLto 55.7639 ng/mL 75.0543 ng · hr/mL 82.1136 ng · hr/mL Estradiol 12.9580pg/mL to 280.7467 pg · hr/mL to 491.8879 pg · hr/mL to 20.2469 pg/mL438.6667 pg · hr/mL 768.5748 pg · hr/mL Free estrone 85.3098 pg/mL to1819.2181 pg · hr/mL to 2398.9939 pg · hr/mL to 133.2966 pg/mL 2842.5283pg · hr/mL 3748.4280 pg · hr/mL Total estrone 7.0858 ng/mL to 40.3505 ng· hr/mL to 42.6480 ng · hr/mL to 11.0715 ng/mL 63.0476 ng · hr/mL66.6376 ng · hr/mL

TABLE 21 pK Ranges for the Pharmaceutical Composition of Table 4(Pharmaceutical Composition D) C_(max) AUC_((0-t)) AUC_((0-∞))Progesterone 35.6889 ng/mL to 48.0348 ng · hr/mL to 52.5527 ng · hr/mLto 55.7639 ng/mL 75.0543 ng · hr/mL 82.1136 ng · hr/mL Estradiol 25.9161pg/mL to 561.4933 pg · hr/mL to 983.7758 pg · hr/mL to 40.4939 pg/mL877.3333 pg · hr/mL 1537.1496 pg · hr/mL Free estrone 170.6197 pg/mL to3638.4363 pg · hr/mL to 4797.9878 pg · hr/mL to 266.5933 pg/mL 5685.0567pg · hr/mL 7496.8559 pg · hr/mL Total estrone 14.1716 ng/mL to 80.7010ng · hr/mL to 85.2961 ng · hr/mL to 22.1431 ng/mL 126.0953 ng · hr/mL133.2751 ng · hr/mL

It will be apparent to those skilled in the art that variousmodifications and variations can be made in the present disclosurewithout departing from the spirit or scope of the disclosure. Thus, itis intended that the present disclosure cover the modifications andvariations of this disclosure provided they come within the scope of theappended claims and their equivalents.

Likewise, numerous characteristics and advantages have been set forth inthe preceding description, including various alternatives together withdetails of the structure and function of the devices or methods. Thisdisclosure is intended as illustrative only and as such is not intendedto be exhaustive. It will be evident to those skilled in the art thatvarious modifications may be made, especially in matters of structure,materials, elements, components, shape, size and arrangement of partsincluding combinations within the principles of the disclosure, to thefull extent indicated by the broad general meaning of the terms in whichthe appended claims are expressed. To the extent that these variousmodifications do not depart from the spirit and scope of the appendedclaims, they are intended to be encompassed therein.

What is claimed is:
 1. A method of treating a female human patienthaving vasomotor symptoms associated with estrogen deficiency, themethod comprising orally administering to the female human patient inneed of treatment thereof with food a soft gelatin capsule comprising afill material, the fill material consisting of: a. a solubilizing agentconsisting of: i. about 196 mg of mono- and diglycerides of caprylic andcapric acid; and ii. about 3 mg of at least one of lauroyl macrogol-32glycerides EP, lauroyl polyoxyl-32 glycerides NF, or lauroylpolyoxylglycerides; b. 1 mg of 17β-estradiol, wherein the 17β-estradiolis fully solubilized in the solubilizing agent; and c. 100 mg ofprogesterone, wherein a first portion of the progesterone is solubilizedin the solubilizing agent and a second portion of the progesterone ismicronized; wherein the second portion of the progesterone is uniformlydispersed in the solubilizing agent; and wherein the administration ofthe capsule to the patient produces one or more pharmacokineticparameters selected from the group consisting of: i. an area under thecurve (AUC)(0-t) for 17β-estradiol ranging from 561.4933 pg·hr/ml to877.3333 pg·hr/ml; ii. a Cmax for 17β-estradiol ranging from 25.9161pg/ml to 40.4939 pg/ml; iii. an AUC(0-t) for estrone ranging from3638.4363 pg·hr/ml to 5685.0567 pg hr/ml; iv. a Cmax for estrone rangingfrom 170.6197 pg/ml to 266.5933 pg/ml; v. an AUC(0-t) for progesteroneranging from 48.0348 ng·hr/ml to 75.0543 ng·hr/ml; and vi. a Cmax forprogesterone ranging from 35.6889 ng/ml to 55.7639 ng/ml.
 2. The methodof claim 1, wherein the patient has a uterus.
 3. The method of claim 1,wherein the administration of the capsule to the patient furtherproduces a Tmax for progesterone that ranges from 2.4 hr to 3.8 hr or aTmax for estrone ranging from 4.4 hr to 6.9 hr.
 4. The method of claim3, wherein the administration of the capsule to the subject producesthree or more of the pharmacokinetic parameters.
 5. The method of claim1, wherein the administration of the capsule to the patient produces thefollowing pharmacokinetic parameters: a. an AUC(0-t) for 17β-estradiolranging from 561.4933 pg·hr/ml to 877.3333 pg·hr/ml; b. a Cmax for17β-estradiol ranging from 25.9161 pg/ml to 40.4939 pg/ml; c. anAUC(0-t) for estrone ranging from 3638.4363 pg·hr/ml to 5685.0567pg·hr/ml; d. a Cmax for estrone ranging from 170.6197 pg/ml to 266.5933pg/ml; and e. an AUC(0-t) for progesterone ranging from 48.0348 ng·hr/mlto 75.0543 ng·hr/ml.
 6. The method of claim 1, wherein at least about15% by weight of the progesterone is solubilized in the solubilizingagent, and at least about 50% by weight of the progesterone ismicronized.
 7. The method of claim 6, wherein at least about 30% byweight of the progesterone is solubilized in the solubilizing agent. 8.The method of claim 1, wherein the micronized progesterone has an X50particle size of below about 15 microns, an X90 particle size of belowabout 25 microns, or both.
 9. A method of treating a female humanpatient having vasomotor symptoms associated with estrogen deficiency,the method comprising orally administering to the female human patientin need of treatment thereof with food a soft gelatin capsule comprisinga fill material, the fill material comprising: a. a solubilizing agentcomprising: i. about 196 mg of mono- and diglycerides of caprylic andcapric acid; and ii. about 3 mg of at least one of lauryl macrogol-32glycerides EP, lauroyl polyoxyl-32 glycerides NF or lauroylpolyoxylglycerides; b. 1 mg of 17β-estradiol, the 17β-estradiol beingfully solubilized in the solubilizing agent; c. 100 mg of progesterone,wherein a first portion of the progesterone is solubilized in thesolubilizing agent and a second portion of the progesterone ismicronized, and wherein at least about 20% by weight of the progesteroneis solubilized in the solubilizing agent; wherein the second portion ofthe progesterone is uniformly dispersed in the solubilizing agent; andwherein the administration of the capsule to the patient produces one ormore pharmacokinetic parameters selected from the group consisting of:i. an area under the curve (AUC)(0-t) for 17β-estradiol ranging from561.4933 pg·hr/ml to 877.3333 pg·hr/ml; ii. a Cmax for 17β-estradiolranging from 25.9161 pg/ml to 40.4939 pg/ml; iii. an AUC(0-t) forestrone ranging from 3638.4363 pg·hr/ml to 5685.0567 pg·hr/ml; iv. aCmax for estrone ranging from 170.6197 pg/ml to 266.5933 pg/ml; v. anAUC(0-t) for progesterone ranging from 48.0348 ng·hr/ml to 75.0543ng·hr/ml; and vi. a Cmax for progesterone ranging from 35.6889 ng/ml to55.7639 ng/ml.
 10. The method of claim 9, wherein the patient has auterus.
 11. The method of claim 9, wherein the administration of thecapsule to the patient further produces a Tmax for progesterone rangingfrom 2.4 hr to 3.8 hr or a Tmax for estrone ranging from 4.4 hr to 6.9hr.
 12. A method of treating a female human patient having vasomotorsymptoms associated with estrogen deficiency, the method comprisingorally administering to the patient in need of treatment thereof withfood a soft gelatin capsule comprising a fill material, the fillmaterial comprising: a. a solubilizing agent comprising: i. a mediumchain oil, wherein at least about 80% of fatty acid chains present inthe oil are C6-C12 and the medium chain oil comprises about 60% to about85% of the weight of the fill material; and ii. a non-ionic surfactant,wherein the non-ionic surfactant comprises about 0.1% to about 10.0% ofthe weight of the fill material; b. 1 mg of 17β-estradiol, the17β-estradiol being fully solubilized in the solubilizing agent; c. 100mg of progesterone, wherein a first portion of the progesterone issolubilized in the solubilizing agent and a second portion of theprogesterone is micronized; wherein the second portion of theprogesterone is uniformly dispersed in the solubilizing agent; andwherein the administration of the capsule to the patient produces one ormore pharmacokinetic parameters selected from the group consisting of:i. an area under the curve (AUC)(0-t) for 17β-estradiol ranging from561.4933 pg·hr/ml to 877.3333 pg·hr/ml; ii. a Cmax for 17β-estradiolranging from 25.9161 pg/ml to 40.4939 pg/ml; iii. an AUC(0-t) forestrone ranging from 3638.4363 pg·hr/ml to 5685.0567 pg·hr/ml; iv. aCmax for estrone ranging from 170.6197 pg/ml to 266.5933 pg/ml; v. anAUC(0-t) for progesterone ranging from 48.0348 ng·hr/ml to 75.0543ng·hr/ml; and vi. a Cmax for progesterone ranging from 35.6889 ng/ml to55.7639 ng/ml.
 13. The method of claim 12, wherein the patient has auterus.
 14. The method of claim 12, wherein the administration of thecapsule to the patient further produces a Tmax for progesterone rangingfrom 2.4 hr to 3.8 hr or a Tmax for estrone ranging from 4.4 hr to 6.9hr.
 15. The method of claim 12, wherein the ratio of the medium chainoil to the non-ionic surfactant is between about 60:1 to about 70:1. 16.The method of claim 12, wherein at least about 20% by weight of theprogesterone is solubilized in the solubilizing agent.
 17. A method oftreating a female human patient having vasomotor symptoms associatedwith estrogen deficiency, the method comprising orally administering tothe patient in need of treatment thereof with food a soft gelatincapsule comprising a fill material, the fill material comprising: a. asolubilizing agent; b. 1 mg of 17β-estradiol, the 17β-estradiol beingfully solubilized in the solubilizing agent, wherein the 17β-estradiolcomprises about 0.15% to about 0.4% by weight of the fill material; c.100 mg of progesterone, wherein a first portion of the progesterone issolubilized in the solubilizing agent and a second portion of theprogesterone is micronized, wherein at least about 15% by weight of theprogesterone is solubilized in the solubilizing agent, and wherein theprogesterone comprises about 30% to about 35% by weight of the fillmaterial; and wherein the second portion of the progesterone isuniformly dispersed in the solubilizing agent; and wherein theadministration of the capsule to the patient produces one or morepharmacokinetic parameters selected from the group consisting of: i. anarea under the curve (AUC)(0-t) for 17β-estradiol ranging from 561.4933pg·hr/ml to 877.3333 pg·hr/ml; ii. a Cmax for 17β-estradiol ranging from25.9161 pg/ml to 40.4939 pg/ml; iii. an AUC(0-t) for estrone rangingfrom 3638.4363 pg·hr/ml to 5685.0567 pg hr/ml; iv. a Cmax for estroneranging from 170.6197 pg/ml to 266.5933 pg/ml; v. an AUC(0-t) forprogesterone ranging from 48.0348 ng·hr/ml to 75.0543 ng·hr/ml; and vi.a Cmax for progesterone ranging from 35.6889 ng/ml to 55.7639 ng/ml. 18.The method of claim 17, wherein the patient has a uterus.
 19. The methodof claim 17, wherein the fill material further comprises a surfactant,the surfactant comprising about 0.1% to about 5% by weight of the fillmaterial.
 20. The method of claim 17, wherein the surfactant is an ionicsurfactant, a non-ionic surfactant, or a combination thereof.
 21. Themethod of claim 20, wherein the non-ionic surfactant is a medium chainfatty acid ester of polyethylene glycol.
 22. The method of claim 20,wherein the non-ionic surfactant is a polyethylene glycol glyceridecomposed of mono-, di-, and triglycerides and mono- and diesters ofpolyethylene glycol.
 23. The method of claim 20, wherein the non-ionicsurfactant is PEG-32 glyceryl laurate EP.
 24. The method of claim 20,wherein the ratio of the solubilizing agent to the non-ionic surfactantis about 65:1.
 25. The method of claim 17, wherein the micronizedprogesterone has an X50 particle size of below about 15 microns, an X90particle size of below about 25 microns, or both.
 26. The method ofclaim 17, wherein the solubilizing agent comprises a medium chain oil.27. The method of claim 26, wherein the medium chain oil comprises atleast about 75% by weight of medium chain fatty acids.
 28. The method ofclaim 19, wherein the ratio of the solubilizing agent to the surfactantis about 60:1 to about 70:1.
 29. The method of claim 28, wherein theratio of the solubilizing agent to the surfactant is about 65:1.
 30. Themethod of claim 17, wherein at least about 20% by weight of theprogesterone is solubilized in the solubilizing agent.